Fig. 2

Rapamycin affects Prox1 protein but not its transcript. a HepG2 cells were treated with various concentrations of rapamycin from 1 to 50 nM. After 48 h, cells were harvested and analyzed by western blot. Phosphorylated mTOR (p-mTOR) was detected to confirm the inhibitory effect of rapamycin. mTOR and β-actin were measured as quantitative controls. (*: p < 0.05, ***: p < 0.001). b The expression of Prox1 in HepG2 cells was down-regulated by rapamycin after 24 h and decreased at 48 h. c Total RNAs were prepared from HepG2 cells incubated with or without rapamycin and analyzed by semi-quantitative RT-PCR (right panel) and real-time quantitative RT-PCR (left panel). d HepG2 cells were treated with or without rapamycin for 12 h and exposed to additional incubation with cycloheximide. The expression of Prox1 was maintained at 6 h after cycloheximide treatment in DMSO-treated HepG2 cells, but it decreased after 4 h in rapamycin-treated cells