Figure 4

Anti-HPV and anti-cancer effect of B. pinnata crude leaf extract and fraction F4 in cervical cancer cells. A. Northern blot analysis of HPV18 positive HeLa cells incubated with indicated concentrations of crude leaf extract and fraction F4. Quantity and quality of total RNA (15 μg/lane) extracted was examined on agarose gel by assessing 28S & 18S ribosomal RNAs (upper panels). The membranes were hybridized with HPV18-specific probe (middle panel), stripped and rehybridized with β-actin -specific DNA probe as an internal control to assess equal loading (lower panels). B. Aggregated mean (± S.D.) abundance ratio of HPV18 transcript w.r.t. to β-actin following treatment with indicated doses of crude extract (a) and fraction F4 (b) in three independent experiments. The abundance ratio HPV18 transcript to β-actin mRNA was analyzed by densitometry as described in "Methods" and their ratio in untreated control was used as reference. C. Immunoblotting analysis of Bcl-2, and Bax, caspase-3, poly (ADP-ribose) polymerase-1 (PARP- 1), in fraction F4 treated HeLa cells at different time-intervals. D. The abundance ratio of indicated proteins to β-actin was analyzed by densitometry and their ratio in untreated control was used as reference. The values indicate aggregated mean ± S.D. of three independent experiments. *p < 0.05 versus untreated control cultures.