Figure 2.

H₂O₂ regulated PUMA expression at transcriptional level. (A, B) Dose and time response of LoVo cells treated with H₂O₂. PUMA mRNA expression was detected by RT-PCR using 0.5 μg total RNA. Beta-actin mRNA was amplified as an internal control. (C) LoVo cells were transiently transfected with the -336/+157 PUMA-luc reporter plasmid (0.5 μg). H₂O₂ (0.64 mM) was added to the indicated samples 24 hours post-transfection and exposure was continued for 15 minutes. The pSVβ-Galactosidase plasmid (0.1 μg) was included in each sample for normalization of transfection variability. Luciferase activity was determined in cell lysates 48 hours following the transfection and the values (mean ± SD, *p < 0.05) from at least two independent expression performed in duplicate are shown in the form of a bargraph.