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. 2016 Sep 28;6:34377. doi: 10.1038/srep34377

Figure 3. Effect of cyclopamine on the expression of HPV16 E6 and E7 oncogenes and their downstream targets.

Figure 3

(a) Evaluation of E6 and E7 transcript level in cyclopamine-treated cervical cancer cells by RT-PCR. Representative photograph of EtBr-stained 2% agarose gel showing PCR amplification of E6 and E7 in cDNA prepared from cyclopamine (20 μM) treated and untreated HeLa and SiHa cells. GAPDH was used as input control for normalization. (a, lower panel) Aggregated mean (±S.D.) abundance ratios of the transcripts w.r.t. GAPDH in three independent experiments. *p value ≤ 0.05 versus untreated control cells. (b) Representative immunoblots showing expression of HPV16/18 E6, HPV16/18 E7 and their downstream target, p53 and pRB respectively in cellular proteins (50 μg/lane) from HeLa and SiHa cells treated with cyclopamine (20 μM) for 24 h and evaluated by SDS-PAGE followed by immunoblotting. Blots were striped and reprobed for β-actin as input control. (b, lower panel) Aggregated mean (±S.D.) abundance ratios of the band intensity of E6, E7, p53 and pRb proteins in HeLa and SiHa cells normalized to β-actin in three independent experiments. *p value ≤ 0.05 versus untreated control cells.