Figure 2

Effect of UVC-treatment on endogenous AP-1 activity and cervicosphere formation in CaCxSLC. (A) Status of AP-1 signaling in CaCxSLCs derived from SiHa cells. Representative autoradiogram showing AP-1 specific DNA-binding activity in cells from parental, cervicospheres derived CaCxSLCs and non-CaCxSLCs. Nuclear protein (10 µg/lane) from CaCxSLC cells (DCM) or non-CaCxSLCs cells (CM) were co-incubated with radiolabelled (γ-³²P-ATP) AP-1 consensus sequence that contain AP-1 binding sequence and AP-1 specific DNA-binding was examined (i). AP-1 specific DNA-binding activity was verified by cold competition as described in Method by co-incubating 100x molar excess of either unlabelled homologous AP-1 probe or heterologous Oct-1 probe (ii). Numbers on the bottom represent fold change in AP-1 binding vs. parental cells. Data are expressed as the mean ± SD of three independent experiments (iii). *p-value < 0.05 vs. untreated controls i.e. CaCxSLCs and non-CaCxSLC cells. Untreated parental SiHa cells were included as positive control. (B) Effect of UV on AP-1 DNA-binding activity. Representative autoradiogram showing AP-1 DNA-binding in nuclear proteins (10 µg/lane) derived from cells of indicated culture that were exposed to 100 J/m² UV radiation for variable time interval and harvested after 12 h after completion of the treatment (i). Cumulative quantitative densitometry data of three independent experiments (ii). *p-value < 0.05 vs. untreated cervicospheres of CaCxSLCs, ^# p-value < 0.05 vs untreated non-CaCxSLCs.