Fig 2: hnRNP E1 effects on HPV16 oncogene and subsequent changes in cell biological function. Note: OE indicates hnRNP E1 overexpression group and NC-OE indicates the relative control group. KD indicates hnRNP E1 knockdown group and NC-KD indicates the relative control group. *P < 0.05. (A) The expression of HPV16 E2, and E6 were detected in SiHa cells by RT-qPCR and western blotting. (B) The relationship between HPV16 E6 expression, cell proliferation, and apoptosis in hnRNP E1-modified cells. The expression of HPV16 E2, and E6 were detected in SiHa cells by RT-qPCR and western blotting. The PI and total apoptosis rate were evaluated using flow cytometry analyses. As HPV16 E6 mRNA and protein expression levels decreased, the proliferation indices of SiHa cells decreased, while the total apoptosis rate increased. As HPV16 E6 expression increased, the proliferation indices of SiHa cells increased, while the total apoptosis rate decreased.

Fig 3: hnRNP E1 mediated the inhibition of apoptosis and promotion of autophagy induced by JL014. (A) HUVECs were treated with 5 µM or 10 µM JL014 in basal M199 medium with serum and FGF-2 deprivation for 3h, 6h and 12 h and western blot analysis of hnRNP E1 protein levels; (B) The protein level of hnRNP E1 were calculated based on densitometry analysis; (C) qPCR analysis of hnRNP E1 mRNA levels after JL014 treated HUVECs 6 h; (D-H) HUVECs were treated with 20 nM hnRNP E1 siRNA or scramble siRNA for 36 h and stimulated with 10 µM JL014 for up 12 h and (D) Nuclear fragmentation of cells by Hoechst 33258 and TUNEL staining (10×). Immunostaining of LC3 puncta (20×); (E) Bar chart showing quantification of HUVEC apoptosis percentage according to Hoechst 33258 staining; (F) Bar chart showing quantification of LC3 puncta per cell; (G) Western blot analysis of LC3 protein level; (H) The protein level of LC3-II were calculated based on densitometry analysis. Data are means ± SEM. *P < 0.05 and **P < 0.01. n=3.

Fig 4: Effects of hnRNP E1 on cell biological function in cervical cancer cell lines. Note: OE indicates hnRNP E1 overexpression group and NC-OE indicates the relative control group. KD indicates hnRNP E1 knockdown group and NC-KD indicates the relative control group. *P < 0.05. (A) Stably transfected hnRNP E1-modified cervical cancer cells were identified by RT-qPCR and western blotting. (B) hnRNP E1 overexpression strongly suppressed cell viabilities in SiHa and C33A cells. hnRNP E1 knockdown markedly enhanced cell viabilities in SiHa and C33A cells. (C) hnRNP E1 arrests the cell cycle from the G0/G1 to the S phase. (D) hnRNP E1 promotes cell apoptosis.

Fig 5: hnRNP E1 expression and correlations with HPV16 E2 and E6 in cervical lesions. (A) hnRNP E1 protein expression levels in the clinical samples. (B) The hnRNP E1 mRNA expression levels in NC and SCC samples, data were obtained from GEO dataset. (C) The hnRNP E1 protein expression levels in HPV+ and HPV- samples. (D) The hnRNP E1 mRNA expression levels in HPV+ and HPV- samples, data were available from GEO dataset. (E) hnRNP E1, HPV16 E2 and E6 expression levels was detected using western blotting in cervix tissues. ß-actin was used as loading control. (F) Correlations between hnRNP E1, HPV16 E2 and E6 in cervical lesions. a/b/c/d, different letters indicate significant differences at least P