Priority Research Paper Volume 1, Issue 6 pp 515—528

Inhibition of mammalian S6 kinase by resveratrol suppresses autophagy

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Figure 2. Resveratrol suppresses autophagy under TOR inhibition. (A) HEK293 cells stably expressing GFP-LC3 growing in complete media were pretreated with DMSO or 50 μM resveratrol (Res) for 1 hour, prior to addition of DMSO or 200 nM rapamycin for 4 hours. 40X magnification fields have been cropped and zoomed for ease of punctae visualization. (B) Quantification of punctae/cell from (A) of 10 fields per treatment are represented as a percentage of DMSO treated cells. Error bars represent s.d.m. * (p < 0.0001) (C) HEK293 GFP-LC3 cells were pretreated for 1 hour with DMSO or resveratrol and subsequently treated with DMSO or 1 mM rapamycin in the presence or absence of 100 nM Bafilomycin A1 for 4 hours. A representative western blot of endogenous LC3 and tubulin are shown. Numbers represent the ratio of LC3-II to tubulin for each condition normalized to Rapamycin in the absence of BafA1.