Anti-Zfp281/ZNF281 antibody

• WB

Unknown

Western blot - Anti-Zfp281/ZNF281 antibody (AB101318)

All lanes:

Western blot - Anti-Zfp281/ZNF281 antibody (ab101318) at 1 µg/mL

Lane 1:

wild type mouse embryonic stem cell lysate at 10 µg

Lane 2:

Zfp281/ZNF281 null mouse embryonic stem cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 97 kDa

Observed band size: 120 kDa,78 kDa

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Exposure time: 4min

• WB

CiteAb

Western blot - Anti-Zfp281/ZNF281 antibody (AB101318)

Zfp281/ZNF281 western blot using anti-Zfp281/ZNF281 antibody ab101318. Publication image and figure legend from Huang, X., Balmer, S., et al., 2017, Elife, PubMed 29168693.

ab101318 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab101318 please see the product overview.

Zfp281 cooperates with Oct4 and P300 for regulation of genes encoding Nodal pathway components.(A) Protein expression of p-Smad2 and Smad2 in WT (J1, CJ7, WT clone no 3), Zfp281KO (clone no. 2.6, 7, 3.34) ESC and WT EpiSC (OEC2, EpiSC9) cells. P-Smad2/Smad2 density ratios were quantified on the right panel. Student T-test was used examine statistical significance, *p<0.05. (B) Co-IP of Zfp281 with Oct4 and P300 in EpiSCs. The non-specific band is IgG heavy chain. (C) ChIP-seq tracks of Zfp281, Oct4 and P300 in ESCs and EpiSCs/EpiLCs at Nodal, Lefty2, and Foxh1 loci. Intensities are shown as mapped reads per million total mapped (RPM). (D) ChIP-qPCR of Zfp281 at Nodal, Lefty2, and Foxh1 regulatory regions in in ESCs and EpiSCs. (E) ChIP-qPCR of histone markers H3K27me at Nodal, Lefty2, and Foxh1 regulatory regions in ESCs and EpiSCs. (F) Diagram of ChIP experiments on E6.5 (WT) embryos. (G) ChIP-qPCR of Zfp281 at Nodal, Lefty2, and Foxh1 regulatory regions in E6.5 (WT) embryos. ChIP experiments were performed in two independent replicates. ChIP-qPCR primer sequences are provided in Figure 7—source data 1.10.7554/eLife.33333.029Figure 7—source data 1.ChIP-qPCR primer sequences.ChIP-qPCR primer sequences.Zfp281 is required to maintain Lefty expression in vitro.(A) Protein expression of p-Smad2, Smad2, Lefty (an antibody detecting both Lefty1 and Lefty2) by two Zfp281 shRNAs knockdown in EpiSCs. P-Smad2/Smad2 and Lefty/Gapdh density ratios were quantified on the bottom panels. (B) Protein expression of p-Smad2, Smad2, Lefty (an antibody detecting both Lefty1 and Lefty2) by treatment of ALK inhibitor (ALKi) in ESCs (CJ7, WT clone no 3) and EpiSCs (OEC2, EpiSC9). P-Smad2/Smad2 and Lefty/Gapdh density ratios were quantified on the bottom panels. Student T-test was used examine statistical significance : ns = non significant, *p<0.05, **p<0.01, ***p<0.001.Zfp281 is required for ESCs differentiating to primitive streak (PS)-like cells.(A) Phase images of ESCs and PS-like like cells with control, ALK inhibitor (ALKi) or Zfp281 shRNA treatments. (B) RT-qPCR for expression of T and Lefty2 at different days in ESC-to-PS-like cells differentiation with vehicle or ALKi treatment. (C) RT-qPCR for expression of T and Lefty2 at different days in ESC-to-PS-like cells differentiation with empty vector or Zfp281 shRNA treatment.Zfp281, Oct4, and P300 colocalize in ESCs and EpiSCs/EpiLCs.ChIP-seq profiles of Otx2, p300, and Oct4 (Buecker et al., 2014) at Zfp281 peak regions in ESCs and EpiLCs. Intensity is plotted within 5 kb around Zfp281 peak center in ESCs and EpiSCs, respectively.

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