Figure 3

Effect of B. pinnata crude leaf extract on AP1 specific DNA-binding and expression of its constituent subunits in cervical cancer cells. A. Nuclear proteins (10 μg/lane) of HeLa cells treated with indicated concentrations of B. pinnata crude extracts (a) or its fraction F4 (b) for 24 h and were assayed for AP1 DNA binding activity by EMSA. Untreated HeLa nuclear proteins and labeled AP1 probe were incubated in the presence of 100 fold molar excess of unlabeled homologous AP1 probe or heterologous Oct-1 probe and assayed for AP1 specific binding by EMSA (c). Fold change was calculated following densitometric evaluation of shifted band. B. Effect of B. pinnata leaf extracts on the expression of AP1 family proteins. Representative immunoblots of cellular proteins isolated from HeLa cells treated with B. pinnata crude leaf extract (a) and fraction F4 (b) as indicated showing expression of different members of AP1 family. β-actin served as a loading control. C. Aggregated mean (± S.D.) abundance ratios of indicated AP1 proteins w.r.t. to β-actin following treatment with indicated doses of crude extract and fraction F4 in three independent experiments. Band intensity of AP1 proteins and β-actin was measured by densitometry as described in "Methods" and their ratio in untreated control was used as reference. *p < 0.05 versus untreated control cultures.