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. 2021 Apr 7;41(4):BSR20203488. doi: 10.1042/BSR20203488

Figure 2. The depletion of eIF4A3 increases HPV16 E7 protein expression.

Figure 2

(A) Western blotting analyses showing endogenous E6 (left panel), E7 (right panel) and eIF4A3 protein levels in CTL or eIF4A3-depleted SiHa cells. β-actin was used as a loading control. (B) Representative scheme of unspliced and alternatively spliced HPV16 early transcripts. Real-time PCR primer sets are in dotted lines (E6All, E6, E6*I and E7) and RT-PCR primer sets are represented by arrows (F6 and R7). (C) RT-PCR showing alternative splicing patterns of HPV16 E6/E7 transcripts in CTL or eIF4A3-depleted SiHa cells. KPNA1 was used as a positive control of eIF4A3 depletion efficiency. B2M and 18S were both used as loading controls. (D) RT-qPCR analysis of HPV16 E6all, E6, E6*I and E7 expression in SiHa cells treated with siRNA-CTL or siRNA-eIF4A3 for 48 h. 18S was used for normalization. Data are represented as means of seven independent experiments. Error bars = s.e.m.; **P<0.01; ***P<0.001; two-tailed Student’s t test. (E) E6/E6All ratio of mRNA levels quantified by real-time PCR in CTL or eIF4A3-depleted SiHa cells. Data are represented as means of seven independent experiments. Error bars = s.e.m.