Fig. 1

dsRNAs targeting the p21 gene promoter induce p21 expression in different human cell lines. Cells were transfected with 50 nM dsRNA for 72 h. The mRNA and protein levels were analyzed using RT-qPCR and Western blotting, respectively. a A schematic representation of the p21 promoter with its transcriptional start site and the dsRNA target. b p21 and 18S rRNA expression levels were assessed using RT-qPCR in PC-3, T-24, and ACHN cells after dsP21-322, dsControl, or mock transfections. The p21 expression level was normalized to that of 18S rRNA and plotted as fold change relative to the mock treatment. The results are expressed as the mean ± S.D. of three independent experiments. c The induction of p21 protein expression was confirmed by Western blot analysis in PC-3, T-24, and ACHN cells. GAPDH levels were also detected and served as a loading control. d p21 mRNA expression was analyzed using RT-qPCR, and the results were normalized to 18S RNA in U2-OS, HeLa, Hep3B, and 293T cells after dsP21-322, dsControl, or mock transfections. The results are expressed as the mean ± S.D. for three independent experiments. e Western blot analysis of p21 and GAPDH after dsP21-322, dsControl, or mock transfections in U2-OS, HeLa, Hep3B, and 293T cells