Fig. 2.

Validation of genes modulated by Bcl-2, enriched by KEGG and associated to Hippo pathway. A, D qRT-PCR analysis of NF2, TEAD4, TP73, CCND3, TEAD2, BIRC5, MYC and SOX2 mRNA expression in A375 (A) and MDA-MB-231 (D) cells transiently transfected with siRNA SMARTpools targeting Bcl-2 (si-Bcl-2) or with control siRNA (si-Ctrl). si-Bcl-2 respect to si-Ctrl is reported. B Western blot analysis of Bcl-2 protein expression and C qRT-PCR analysis of Bcl-2 mRNA level in MDA-MB-231. Representative images of three independent experiments with similar results. HSP72/73 was used to check equal loading and transfer. A, C, D Data represent ratio (mean ± standard error of mean) of mRNA expression normalized to β-actin gene in silenced cell condition versus controls from three independent biological repeats (n = 3), except for BIRC5 and SOX2 in A (n = 5) and TEAD4 in D (n = 4). Statistical analysis was performed applying unpaired two-tailed student’s t test *p < 0.05, **p < 0.01, ***p < 0.001