Anti-VASP antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VASP antibody (AB229624)

HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for VASP (green) using ab229624 at a 1/500 dilution in ICC/IF. Cells were fixed in 4% paraformaldehyde at RT for 10 minutes.

Nuclear counterstain : Hoechst 33342 (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VASP antibody (AB229624)

Paraffin-embedded MDA-MB-231 xenograft stained for VASP with ab229624 at 1/500 dilution in immunohistochemical analysis.

• WB

CiteAb

Western blot - Anti-VASP antibody (AB229624)

Western Blotting using Anti-VASP antibody, ab229624. Publication image from Dang, Y. et al., 2020, Theranostics, 32206125. Legend direct from paper.

PDPK1 and VASP are direct transcriptional targets of HOXC10. (A) Western blotting analysis of PDPK1 and VASP expression in the indicated HCC cells. (B) Real-time PCR analysis of PDPK1 and VASP expression in the indicated HCC cells. (C) HOXC10 transactivates PDPK1 and VASP promoters. The PDPK1 or VASP promoter luciferase construct was cotransfected with pCMV-HOXC10, and promoter activities were detected using a luciferase reporter assay. (D-E) Deletion and selective mutation analyses identified HOXC10-responsive regions in the (D) PDPK1 and (E) VASP promoter. Serially truncated and mutated PDPK1 or VASP promoter constructs were cotransfected with pCMV-HOXC10, and relative luciferase activities were determined. The schematic constructs are shown (left), and the bar graphs present the relative levels of luciferase activity in each of the samples (right). (F-G) ChIP assays demonstrated the direct binding of HOXC10 to the PDPK1 (F) or VASP (G) promoter in Hep3B-HOXC10 cells (left panel) and the enriched binding of endogenous HOXC10 to the PDPK1 or VASP promoter in primary HCC tissues (right panel). Real-time PCR was performed to detect the amounts of immunoprecipitated products. Hepatocytes were separated from the liver tissues of HCC patients and healthy controls (HC). The cells were crosslinked, and the chromatin was immunoprecipitated by anti-HOXC10 or control antibody. All the data are shown as the mean±s.d. * P<0.05 ** P˂0.01.

false

• WB

CiteAb

Western blot - Anti-VASP antibody (AB229624)

Western Blotting using Anti-VASP antibody, ab229624. Publication image from Dang, Y. et al., 2020, Theranostics, 32206125. Legend direct from paper.

HOXC10 is essential for IL-1β-mediated HCC metastasis expression. (A) Hep3B-IL-1β cells were infected with LV-shcontrol or LV-shHOXC10 by lentiviral transduction, and HOXC10 expression was examined by Western blotting. The IL-1β levels in the supernatant of the indicated cells were detected by enzyme-linked immunosorbent assay (ELISA). (B) Transwell assays showed that HOXC10 knockdown inhibited the migration and invasion abilities of Hep3B-IL-1β cells. (C-F) Knockdown of HOXC10 inhibited IL-1β-mediated HCC metastasis. (C) Bioluminescence images, metastasis incidence, and number of lung metastasis foci of the indicated groups of nude mice are shown. (D) Bioluminescence signals. (E) Overall survival. (F) Representative HE staining of lung tissues from the different groups is shown. The scale bars represent 1 mm (low magnification) and 100 µm (high magnification). (G) After Hep3B-IL-1β cells were treated with Anakinra (10 µg/ml) for 24 hr, the protein levels of HOXC10, PDPK1 and VASP were detected by Western blotting. (H) Anakinra treatment (10 µg/ml, 24 hr) significantly inhibited the migration and invasion abilities of Hep3B-IL-1β cells. (I-K) Anakinra treatment markedly inhibited IL-1β-mediated HCC metastasis. (I) Anakinra, 1 mg/kg/day, or PBS, was administered intraperitoneally for 9 weeks. starting 1 week after orthotopic implantation of the tumor. (J) The bioluminescent signals, numbers of lung metastatic foci and incidence of lung metastasis. (K) The overall survival times and representative HE staining of lung tissues from the different groups are shown. The scale bars represent 1 mm (low magnification) and 100 µm (high magnification). (L) A schematic diagram of the role of IL-1β-HOXC10 signaling in inflammation-related HCC metastasis. IL-1β-IL-1R1 signaling upregulates HOXC10 expression through the JNK/c-Jun signaling pathway. PDPK1 and VASP are direct transcriptional targets of HOXC10. HOXC10 promotes HCC invasion and metastasis by upregulating PDPK1 and VASP expression. The IL-1R1 antagonist Anakinra inhibits IL-1β-mediated HOXC10 upregulation, thereby inhibiting IL-1β-HOXC10 signaling-mediated HCC invasion and metastasis.

false