Fig. 5: Effect of in vivo Tfeb KO on mitophagy in β-cells.
a Colocalization of LC3 puncta with TOM20, a mitochondrial marker, in pancreatic islets of Tfeb∆β-cell and TfebF/F mice fed NCD or HFD for 12 weeks estimated by Pearson’s correlation analysis (right). Representative fluorescence images are presented (left). (scale bar, 10 μm) (n = 6) b Colocalization of LAMP2 spot, a lysosomal marker, with TOM20 in pancreatic islets of Tfeb∆β-cell and TfebF/F mice fed NCD or HFD for 12 weeks estimated by Pearson’s correlation analysis (right). Representative fluorescence images are presented (left). (scale bar, 10 μm) (n = 6) c ROS accumulation determined by HNE staining of pancreatic sections from Tfeb∆β-cell and TfebF/F mice fed NCF or HFD for 12 weeks (right). Representative fluorescence images are presented (left). (scale bar, 50 μm) (n = 7 for Tfeb∆β-cell:HFD; n = 8 for Tfeb∆β-cell:NCD; n = 11 for TfebF/F:NCD; n = 14 for TfebF/F:HFD) d Mitochondrial COX activity in pancreatic islets of Tfeb∆β-cell and TfebF/F mice fed NCD or HFD for 12 weeks (right). Representative DAB images are presented (left). (scale bar, 100 μm) (n = 6) e Seahorse XF respirometry using primary pancreatic islets from Tfeb∆β-cell and TfebF/F mice on NCD or HFD for 12 weeks (left). Basal, glucose-stimulated, ATP-coupled and maximal O2 consumptions were calculated from the curves (right). (OCR, O2 consumption rate; FCCP, carbonyl cyanide-4-[trifluoromethoxy]phenylhydrazone; Rot, rotenone; AA, antimycin A) Cells in the rectangles were magnified. All data in this figure are the means ± SEM from more than 3 independent experiments. P values were determined using one-way ANOVA with Tukey’s test; ns, not significant.
