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. Author manuscript; available in PMC: 2015 Apr 23.
Published in final edited form as: Oncogene. 2013 Oct 28;33(43):5100–5108. doi: 10.1038/onc.2013.447

Figure 4. Prohibitin and BASP1 recruit BRG1 to promoters of WT1 target genes.

Figure 4

(A) 48hr after transfection with prohibitin siRNA or control siRNA, U2OS cells were subjected to ChIP analysis with control IgG, anti-BASP1 or anti-BRG1 antibodies. Data are presented as fold enrichment of AREG and c-myc promoters relative to a control genome region. Error bar values are SDM for three independent experiments; *p<0.05 by Student's t test. Immunoblotting with anti-prohibitin or anti-tubulin antibodies confirmed successful depletion of prohibitin. (B) MCF7 cells were subjected to coimmunofluorescence with sheep BASP1 antibodies and rabbit BRG1 antibodies. Hoechst was used to label the nucleus. Control anti-sheep (sh) and control anti-rabbit (r) are shown below. (C) Immunoprecipitation with control IgG antibodies or BASP1 antibodies was performed in HeLa cell nuclear extracts. The samples were then immunoblotted with BASP1 or BRG1 antibodies. I is input. (D) Immunoprecipitation with control IgG antibodies or BRG1 antibodies was performed in HeLa cell nuclear extracts and the samples were immunoblotted with either BRG1 or BASP1 antibodies. I is the input nuclear extract. (E) U2OS cells were transfected with control siRNA or BASP1 siRNA and 48 hr later were subjected to ChIP with control IgG antibodies or BRG1 antibodies. The results are presented as fold enrichments at AREG and c-myc promoters relative to a control genome region. Error bars are obtained from the SDM of three independent experiments (*p<0.05 by Student's t test). Immunoblotting with BASP1 antibodies or tubulin antibodies confirmed successful depletion of BASP1. (F) Vector control K562 cells (V-K562) and BASP1-K562 cells were transfected with control siRNA or BRG1 siRNA and 48 hr later RNA and whole cell extracts were obtained. Samples were immunoblotted with anti-BRG1 or anti-tubulin antibodies to confirm depletion of BRG1. The expression of AREG, Ets-1 and VDR are presented as relative to GAPDH mRNA. Error bars are the SDM of three independent experiments and Student's t test of p<0.05 was performed.