Figure 6.

ARHI expression induces Rab7 and is mediated through FOXo3a. (a) Cell lysates from SKOv3-ARHI cells were examined for Rab7 expression at indicated intervals after treatment with DOX. (b) Cells were treated as in a and the level of Rab7 mRNA was quantified by real-time RT-PCR. The columns indicate the mean, and the bars indicate the S.E. (*P<0.05; **P<0.01). Data were obtained from three independent experiments. (c) SKOv3-ARHI cells were transfected with siControl (C) or siFOXo3a (F) and treated with or without DOX. Rab7 expression was determined by RT-PCR and (d) western blot analysis. (e) ARHI expression induces autophagosome formation. SKOv3-ARHI cells were transfected with mCherry-GFP-LC3 for 24. Bafilomycin-a₁ (BA, 100 nM) was added to the plate for the last 16 h of transfection. Scale bars: 10 μm. (f) A schematic diagram of mCherry-GFP-LC3 degradation. The double-tagged LC3 protein (mCherry-GFP-LC3) will emit yellow (green merged with red) fluorescence in non-acidic structures and appear as red only in the autolysosome owing to quenching of GFP in these acidic structures, (g) ARHI increase LC3-II and decreases p62. Western bot analysis of LC3 and p62 were examined