Fig. 5: ZnT8 LOF enhances SC-β cell survival by alleviating ER stress. | Nature Communications

Fig. 5: ZnT8 LOF enhances SC-β cell survival by alleviating ER stress.

From: ZnT8 loss-of-function accelerates functional maturation of hESC-derived β cells and resists metabolic stress in diabetes

Fig. 5

a qRT-PCR analysis of the expression of ER stress-related genes for WT and KO-1 SC-β cells in the presence of PA (n = 12). b Protein expression of WT and KO-1 SC-β cells treated with PA was determined by western blot analysis (n = 3 independent experiments). c, d Representative immunofluorescent images (c) and mean intensity measurement (d) of sXBP1 in the INS+ cells with PA or solvent control treatment (n = 30 from three independent experiments). e Representative trace of Fluo-4 relative fluorescence intensity (F1/F0) versus time in a single SC-β cell upon PA stimulation. f Maximal peak increase of relative fluorescence intensity upon PA stimulation over an average of baseline fluorescence intensity (0-90 s) (HUES8-WT, n = 58; HUES8-KO, n = 77 from four independent experiments). g Mitochondrial membrane potential measured by JC-10 (mean fluorescence of monomer/aggregate). The samples were treated with PA for 0 (n = 4), 30 min (n = 6), 45 min (n = 7), and 60 min (n = 7). h, i Representative images (h) and mean intensity measurement (i) of sXBP1 in the INS+ cells of HUES8-WT infected with Ad-CT (adenovirus-control) or Ad-ZnT8 (adenovirus-ZnT8) in the presence of PA (n = 30 cells from three independent experiments for each condition). j Representative FACS plots and quantifications of the percentages of Annexin V+/INS-GFP+ cells in the WT SC-β cells treated with PA and infected with adenovirus-ZnT8 or adenovirus-control (n = 7). k Representative FACS plots and quantifications of the Annexin V+/INS-GFP+ percentages in the SC-β cells in the absence or the presence of ZnSO4 (100 μM) (n = 5). Two-way ANOVA with Sidak’s multiple-comparisons was used to analyze for a, d and g. Unpaired two-tailed t tests were used to analyze for f, i, j and k. Scale bars of c and h are 10 μm (low magnification) and 5 μm (high magnification). Data are presented as mean ± s.e.m. Individual data points are shown for all bar graphs. Source data are provided as a Source Data file.

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