Recombinant Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1535(2)] to Androgen Receptor
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Androgen Receptor antibody [EPR1535(2)]
See all Androgen Receptor primary antibodies -
Description
Rabbit monoclonal [EPR1535(2)] to Androgen Receptor -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
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Positive control
- WB: T47-D, LnCaP and 22Rv1 cell lysates; Rat and mouse prostate lysates. IHC-P: Human prostate, prostatic adenocarcinoma, prostatic hyperplasia tissues; Rat and mouse testis tissues; Breast carcinoma and prostatic carcinoma T3 tissues. ICC/IF: MCF7 cells;EP156T-AR, 957E/hTERT-AR and LNCaP cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1535(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Androgen Receptor antibody [EPR1535(2)] (ab194194)
- Alexa Fluor® 647 Anti-Androgen Receptor antibody [EPR1535(2)] (ab194195)
- HRP Anti-Androgen Receptor antibody [EPR1535(2)] (ab194196)
- Anti-Androgen Receptor antibody [EPR1535(2)] - Low endotoxin, Azide free (ab209969)
- Anti-Androgen Receptor antibody [EPR1535(2)] - BSA and Azide free (ab271891)
- Alexa Fluor® 555 Anti-Androgen Receptor antibody [EPR1535(2)] (ab275124)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133273 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (2) |
1/2000. Predicted molecular weight: 98 kDa.
For unpurified use at 1:1000 For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy). |
IHC-P | (7) |
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/100-1/250 |
ICC/IF | (1) |
1/100 - 1/250.
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Notes |
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WB
1/2000. Predicted molecular weight: 98 kDa. For unpurified use at 1:1000 For Lysate preparation protocol, please refer to the protocol book in the protocol section and/or here (downloadable copy). |
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100-1/250 |
ICC/IF
1/100 - 1/250. |
Target
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Function
Steroid hormone receptors are ligand-activated transcription factors that regulate eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Transcription factor activity is modulated by bound coactivator and corepressor proteins. Transcription activation is down-regulated by NR0B2. Activated, but not phosphorylated, by HIPK3 and ZIPK/DAPK3.
Isoform 3 and isoform 4 lack the C-terminal ligand-binding domain and may therefore constitutively activate the transcription of a specific set of genes independently of steroid hormones. -
Tissue specificity
Isoform 2 is mainly expressed in heart and skeletal muscle (PubMed:15634333). Isoform 3 is expressed by basal and stromal cells of prostate (at protein level) (PubMed:19244107). -
Involvement in disease
Androgen insensitivity syndrome
Spinal and bulbar muscular atrophy X-linked 1
Defects in AR may play a role in metastatic prostate cancer. The mutated receptor stimulates prostate growth and metastases development despite of androgen ablation. This treatment can reduce primary and metastatic lesions probably by inducing apoptosis of tumor cells when they express the wild-type receptor.
Androgen insensitivity, partial -
Sequence similarities
Belongs to the nuclear hormone receptor family. NR3 subfamily.
Contains 1 nuclear receptor DNA-binding domain. -
Domain
Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain. In the presence of bound steroid the ligand-binding domain interacts with the N-terminal modulating domain, and thereby activates AR transcription factor activity. Agonist binding is required for dimerization and binding to target DNA. The transcription factor activity of the complex formed by ligand-activated AR and DNA is modulated by interactions with coactivator and corepressor proteins. Interaction with RANBP9 is mediated by both the N-terminal domain and the DNA-binding domain. Interaction with EFCAB6/DJBP is mediated by the DNA-binding domain. -
Post-translational
modificationsSumoylated on Lys-388 (major) and Lys-521. Ubiquitinated. Deubiquitinated by USP26. 'Lys-6' and 'Lys-27'-linked polyubiquitination by RNF6 modulates AR transcriptional activity and specificity.
Phosphorylated in prostate cancer cells in response to several growth factors including EGF. Phosphorylation is induced by c-Src kinase (CSK). Tyr-535 is one of the major phosphorylation sites and an increase in phosphorylation and Src kinase activity is associated with prostate cancer progression. Phosphorylation by TNK2 enhances the DNA-binding and transcriptional activity and may be responsible for androgen-independent progression of prostate cancer. Phosphorylation at Ser-83 by CDK9 regulates AR promoter selectivity and cell growth. Phosphorylation by PAK6 leads to AR-mediated transcription inhibition.
Palmitoylated by ZDHHC7 and ZDHHC21. Palmitoylation is required for plasma membrane targeting and for rapid intracellular signaling via ERK and AKT kinases and cAMP generation. -
Cellular localization
Nucleus. Cytoplasm. Predominantly cytoplasmic in unligated form but translocates to the nucleus upon ligand-binding. Can also translocate to the nucleus in unligated form in the presence of RACK1. - Information by UniProt
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Database links
- Entrez Gene: 367 Human
- Entrez Gene: 11835 Mouse
- Entrez Gene: 24208 Rat
- Omim: 313700 Human
- SwissProt: P10275 Human
- SwissProt: P19091 Mouse
- SwissProt: P15207 Rat
- Unigene: 76704 Human
see all -
Form
There are 2 isoforms produced by alternative splicing. Isoform 1 is also known as: AR-B; isoform 2 is known as AR-A or variant AR45. -
Alternative names
- AIS antibody
- ANDR_HUMAN antibody
- Androgen nuclear receptor variant 2 antibody
see all
Images
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All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/1000 dilution
Lane 1 : Mouse testis lysate
Lane 2 : Rat testis lysate
Lane 3 : Mouse liver lysate
Lane 4 : Rat liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsBlocking/Diluting buffer: 5% NFDM/TBST
Loading Control: Rabbit monoclonal [EPR16891] to GAPDH (ab181602)
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Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/5000 dilution (purified) + Mouse prostate lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 98 kDaBlocking and diluting buffer: 5% NFDM/TBST
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Immunocytochemistry/ Immunofluorescence - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)Image from Azeem W et al., PLoS One. 2017;12(6):e0177861. Fig 6.; doi: 10.1371/journal.pone.0177861. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Texas red (Tx-Red) indirect immunofluorescent detection of Androgen Receptor using ab133273 in (A) EP156T-AR, (B) 957E/hTERT-AR and (C) LNCaP cells. The cells were treated with ± 1 nM R1881 for 24 hours.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)shao et al Oncotarget. 2018 Mar 6; 9(18): 14456–14471. Published online 2018 Feb 12. doi: 10.18632/oncotarget.24470
Formalin-fixed, paraffin-embedded tissues labeling Androgen Receptor with ab133273. Left: Tumor from orthotopic injectionof PTEN KD/TE cells. Right: Human postate cancer tissue.
From Figure 3 of Shao et al/
Shao et al Oncotarget. 2018 Mar 6; 9(18): 14456–14471.Published online 2018 Feb 12. doi: 10.18632/oncotarget.24470
Reproduced under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0).
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Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling Androgen receptor with purified ab133273 at 1:100 dilution (1.3μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Immunohistochemical analysis of paraffin-embedded human prostatic adenocarcinoma tissue labelling Androgen Receptor using unpurified ab133273 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/20000 dilution
Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
Lane 2 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method
Lane 3 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method
Lane 4 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method
Lysates/proteins at 1/15 dilution per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 98 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking/Diluting buffer - 5% NFDM/TBST
The androgen receptor variant band detected in 22RV1 cells is reported by PMID: 22315407.
We recommend you to try both RIPA and 1%SDS Hot lysis preparation methods to get desired bands.
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Different batches of ab133273 were tested on LNCaP (Human prostate carcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 120 kDa.
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Immunofluorescent staining of LnCAP cells labelling Androgen Receptor using unpurified ab133273, at 1/100 dilution
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All lanes : Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/5000 dilution (purified)
Lane 1 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates
Lane 2 : Rat prostate lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 98 kDaBlocking and diluting buffer: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue sections labeling Androgen Receptor with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue sections labeling Androgen Receptor with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue sections labeling Androgen Receptor with Purified ab133273 at 1:500 dilution (0.25 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
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Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273) at 1/1000 dilution (unpurified) + LNCaP (Human prostate carcinoma epithelial cell) at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 98 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking/Diluting buffer 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Immunohistochemical analysis of paraffin-embedded human prostate tissue labelling Androgen Receptor using unpurified ab133273, at1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing positive staining in Breast carcinoma tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing negative staining in Normal brain tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing negative staining in Normal tonsil tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing positive staining in Endometrial carcinoma tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing negative staining in Normal breast tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing negative staining in Normal colon tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing negative staining in Ovarian carcinoma tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing negative staining in Colonic adenocarcinoma tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing negative staining in Normal liver tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Androgen Receptor antibody [EPR1535(2)] (ab133273)
Unpurified ab133273 showing positive staining in Prostatic carcinoma T3 tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (99)
ab133273 has been referenced in 99 publications.
- Yun SI et al. Weekly treatment with SAMiRNA targeting the androgen receptor ameliorates androgenetic alopecia. Sci Rep 12:1607 (2022). PubMed: 35102171
- Azeem W et al. Proteasome-Mediated Regulation of GATA2 Expression and Androgen Receptor Transcription in Benign Prostate Epithelial Cells. Biomedicines 10:N/A (2022). PubMed: 35203681
- Limberger T et al. KMT2C methyltransferase domain regulated INK4A expression suppresses prostate cancer metastasis. Mol Cancer 21:89 (2022). PubMed: 35354467
- Han H et al. Mesenchymal and stem-like prostate cancer linked to therapy-induced lineage plasticity and metastasis. Cell Rep 39:110595 (2022). PubMed: 35385726
- Xia L et al. m6A-induced repression of SIAH1 facilitates alternative splicing of androgen receptor variant 7 by regulating CPSF1. Mol Ther Nucleic Acids 28:219-230 (2022). PubMed: 35402071