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. 2012 Mar 19;3(3):299–313. doi: 10.18632/oncotarget.462

Figure 2. BRCA1/p220 silencing triggers BRCA1-IRIS expression in HME cells.

Figure 2

(A) Western blot (right) or RT/qPCR (left) analysis of the fold induction in BRCA1-IRIS protein normalized to actin or mRNA normalized to GAPDH mRNA, respectively in HME cells silenced (for 72h) from control (Luc), BRCA1/p220 and BARD1 and treated or not with cycloheximide during the last 24h. Data represent the means ± SD from triplicate, done three independent times, whereas ** is a p≤0.01. Far right panels show the effects of BRCA1/p220 (upper panels) and BARD1 (lower panels) siRNA on the expression of their cognate protein in HME cells. RT/qPCR analysis (B) or western analysis (C) of BRCA1-IRIS mRNA or protein, respectively in parental, uninducible IRISa and inducible IRISb and IRISc HME cell lines following control or BRCA1/p220 silencing for 24, 48, 72 or 168h. Data in (B) represent the means ± SD from triplicate, done three independent times, whereas * is a p≤0.05 and ** is a p≤0.01. Right panels in (B) show analysis for BRCA1-IRIS overexpression in the different inducible cell lines (upper panels), and the effect of BRCA1/p220 siRNA on the expression of BRCA1/p220 protein at 0, 72 and 168h (lower panels).