Fig. 6. eIF2S2 downregulates p15 and p27 expression in cervical cancer cells. The p15 and p27 expression level was detected in SiHa cell lysate, where cells were (A) overexpressed with p3x-Flag-CMV10-eIF2S2 transfection or (B) knocked down with eIF2S2 siRNA transfection. The mRNA expression of p15 was detected by qRT-PCR (C) with p3x-Flag-CMV10-eIF2S2 transfection. HPRT was used as an internal control. *p<0.001, compared to vector alone. (D) SiHa cells were transiently transfected with eIF2S2 or control vector and incubated with or without TGFβ (5 ng/ml) for 24 h. The p15 promoter activity from transfected cells was measured as described in the Materials and Methods section. (E) SiHa cells were transiently transfected with p3x-Flag-CMV10-eIF2S2 or control vector. Protein lysates underwent Western blot analysis to detect the expression level of Smad7. Flag and eIF2S2 were measured to confirm the transfection efficiency, and actin was used as the loading control. (F) SiHa cells were transfected with control siRNA or eIF2S2 siRNA, and the Smad7 expression was detected by Western blot analysis. All experiments were performed in triplicate.
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