Fig 2: (a) C-33A cells endogenously express Cx32, unlike HeLa or SiHa cells (n=3). (b) The endogenous GJIC of C-33A cells was abolished by gap junction inhibitors 2APB and 18a-GA (n=3). (c) Western blot comparing effectiveness of three siRNAs in reducing expression of Cx32 in C-33A cells. (n=3). (d) Suppression of SN-induced apoptosis by treatment with 2APB or 18a-GA in C-33A cells (n=7). (e) The anti-apoptotic effect of 2APB and 18a-GA in C-33A cells was reversed by Cx32 RNA interference (n=5). Error bar: standard error. *P<0.05; n=1 represents an independent cell culture

Fig 3: (a) Expression of Cx32, Cx26, Cx30 and Cx43 in normal cervix and CaCx samples. Expression of Cx26, Cx30 and Cx43 was decreased in the cancer samples relative to control, while expression of Cx32 was markedly increased. (b) Western blots showing that expression of expression of Cx32 correlated with increased FIGO score. Data are shown for five samples in each category (c) Immunohistochemistry showing that Cx32 and Cx43 in normal cervix tissue were localized to junctional regions. In CaCx cells, Cx32 aberrantly aggregated in para-nuclear cytoplasm and Cx43 was rarely expressed. (Scale bar: 20 µm). *P<0.05

Fig 4: (a) In 30 CaCx specimens, expression of EGFR was highly correlated with expression of Cx32 (r=0.604, P=0.00041). (b) In C-33A cells, when Cx32 expression was inhibited by Cx32 siRNAs, EGFR expression was significantly suppressed (n=3). (c) In HeLa-Cx32 cells, after induced Cx32 expression, EGFR was significantly increased and its downstream effectors p-Erk1/2 and p-Stat3 were upregulated (n=3). (d) In nuclear protein samples, Cx32 expression in CaCx cells (n=10) was higher than that in normal cervix cells (n=5, P=0.0152). Error bar: standard error. *P<0.05; n=1 represents an independent cell culture; r: Pearson correlation coefficient

Fig 5: (a) In HeLa-Cx32 cells, EGFR siRNA sequence 1 effectively inhibited EGFR expression with and without doxycycline treatment (n=3). (b) The anti-apoptotic effect of Cx32 expression was reversed by siRNA suppression of EGFR (n=3). (c and d) The anti-apoptotic effect of Cx32 expression was reversed by EGFR signaling inhibitors erlotinib and afatinib (n=3). Error bar: standard error. *P<0.05; n=1 represents an independent cell culture