Fig. 6.

Knockdown or overexpression of mTOR interferes with the inhibitory activity of resveratrol on hsBAFF-induced inhibition of autophagy and increase of cell proliferation/viability in B cells. Raji cells, infected with lentiviral shRNA to mTOR or GFP (as control), or infected with Ad-mTOR or Ad-LacZ (for control) and infected with/without Ad-GFP-LC3, respectively, were pretreated with 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, F) 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, G) The relative densities for p-S6K1 (Thr389) to S6K1, and ATG5, LC3-II, p62, survivin to β-actin were semi-quantified using NIH image J. (C, H) The number of GFP-LC3 puncta per cell was quantified by GFP-LC3 assay. (D, I) The relative cell proliferation was evaluated by cell counting. (E, J) The relative cell viability was determined by the MTS assay. All data were expressed as mean ± SE (n = 3 for B, G; n = 6 for C-E, H-J). 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, mTOR shRNA group vs GFP shRNA group; ^dp < 0.05, Ad-mTOR group vs Ad-GFP group.