Fig. 8.

Ectopic expression of constitutively active or dominant negative Akt affects the inhibitory activity of resveratrol on hsBAFF-induced inhibition of autophagy and increase of cell proliferation/viability in B cells. Raji cells, infected with Ad-myr-Akt, Ad-dn-Akt, or Ad-LacZ (for control) and infected with/without Ad-GFP-LC3, respectively, were pretreated with/without resveratrol (10 μM) for 1 h, followed by stimulation with/without hsBAFF (2.5 μg/ml) for 12 h (for Western blotting and GFP-LC3 assay) or 48 h (for cell proliferation/viability assay). (A) Total cell lysates were subjected to Western blotting using indicated antibodies. The blots were probed for β-actin as a loading control. Similar results were obtained in at least five independent experiments. (B) The relative densities for p-Akt (Ser473), p-Akt (Thr308), ATG5, LC3-II, p62, survivin to β-actin were semi-quantified using NIH image J. (C) The number of GFP-LC3 puncta per cell was quantified by GFP-LC3 assay. (D) The relative cell proliferation was evaluated by cell counting. (E) The relative cell viability was determined by the MTS assay. All data were expressed as mean ± SE (n = 3 for B; n = 6 for C-E). Using one-way ANOVA or Student’s t-test, ^ap < 0.05, difference vs control group; ^bp < 0.05, difference vs 2.5 μg/ml hsBAFF group; ^cp < 0.05, Ad-myr-Akt group or Ad-dn-Akt group vs Ad-GFP group.