Figure 2
From: Pomegranate activates TFEB to promote autophagy-lysosomal fitness and mitophagy
PE promotes TFEB nuclear localization via dephosphorylation at TFEBS142 to upregulate autophagy transcriptome. (a) Left: Fluorescence images depicting spatial localization of GFP-TFEB in GFP-TFEB SY5Y stable cells treated with DMSO, S− or 300 μg/ml PE for 6 h and 24 h. White arrows indicate cells that contained nuclear-localized GFP-TFEB. Right: Quantification of percentage TFEB nuclear localization. Nuclei were stained with DAPI. At least 100 cells from random fields were analyzed for each condition. (b) Top: Immunoblots of phosphorylated TFEB at Ser142 (P-TFEBS142) and total TFEB (T-TFEB) in GFP-TFEB SY5Y stable cells treated with DMSO, S− or 300 μg/ml PE for 6 h and 24 h. Bottom: Quantification of P-TFEBS142 levels, calculated as fold change against DMSO control. Full-length blots are presented in Supplementary Figure S7b. (c) Analysis of real-time PCR autophagy array in SY5Y cells treated with DMSO or 300 μg/ml PE for 24 h. Gene expression was calculated as fold change relative to DMSO control. ^indicates genes that are part of the CLEAR network. All values are mean + S.E.M (n = 3–12). Differences against DMSO or S− are significant at *p<0.05, **p<0.01, ***p<0.005 and #p < 0.05. Scale bar, 10 μm.
