AS160 knockdown upregulates the CDK inhibitor p21 in multiple cell types. (A) Screening of mRNA levels of AS160, p16, p19, p21, and p27 in 3T3-L1 fibroblasts infected with scrambled (SCR) or AS160-specific (KD) shRNAs, at indicated times post-plating. Data shown are normalized mean ± s.e.m. (n = 3 represents 3 replicated experiments, same blow). (B) Levels of the indicated proteins in 3T3-L1 SCR and KD fibroblasts, determined with triplicated sample loading. Tubulin severed as the loading control. (C) Western blots of MCF7, HeLa, Huh7, and HEK-293 cells transfected with 2 AS160 siRNAs (KD1 and KD2) or a scrambled siRNA (SCR), at 48 and 72 h post-transfection. (D) Protein levels of phosphorylated-Rb and AS160 in 3T3-L1 SCR and KD fibroblasts.