(A). qRT-PCR of RASSF2 and ASK1 expression in control (IRES) or RASSF1A (RF1A) cells. GAPDH was used as internal control. (B). The expressions of ASK1, phospho-ASK1, p38, phospho-p38, Bcl2, caspase 3 and cleaved-caspase 3 were analyzed by Western blotting. (C). The subcellular distribution of cytochrome c was analyzed by Western blot. (D). Apoptosis in RASSF1A cells transfected with either control (scrambled) or ASK1-specific siRNA (siRNA-ASK1) was determined by FACS (left two panels, sub-G1 population) or Western blotting on cleaved casp-3 (right panel). Representative images from three independent experiments are shown.