Effects of F-53 on levels of firefly luciferase (<i>Luc</i>) mRNA and protein in culture medium.

<p>A: <i>Luc</i> mRNA expression was determined by RT-PCR. Total RNA was isolated from HeLa cells transiently transfected with pCMVluc+ and treated with F-53 or DMSO for 20–24 h. The expression of <i>Luc</i> was normalized relative to the abundance of the endogenous <i>hGAPDH</i> transcripts. B and C: Western blotting after SDS-PAGE (B) and native PAGE (C). Protein samples for SDS- or native PAGE and western blot analysis were obtained from HeLa cells that had been transiently transfected with pCMVluc+ and pCMVβ and treated with F-53, PTC124, or DMSO for 20–24 h before lysis. The protein level of Luc was normalized relative to that of β-gal. The asterisks indicate statistically significant differences (* p<0.05, ** p<0.01) between F-53 and DMSO treatments, determined using post-hoc analysis with Fisher's LSD test. D: DMSO, Luc: Luciferase. Each value represents the mean±SEM of three independent experiments.