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. 2022 Aug 8;13:4621. doi: 10.1038/s41467-022-32425-7

Fig. 5. Impairment of lysosome activity correlates with misfolded proinsulin accumulation during chronic ER stress.

Fig. 5

a Scatterplot of fold changes in CPA18 vs CON MIN6 cells, with the repressed gene expression indicated in pink, and the KEGG pathway analysis of the highlighted group, below a. Scatterplots of fold change in acute (CPA1/CON) and chronic (CPA18/CON) ER stress for genes related to the proteasome (b) and the lysosome (c). d lysosome enzymatic activity measured in MIN6 cells treated with Baf A1 or CPA. n = 3 independent experiments. Error bars represent S.E.M. p-value represents the statistical test by two-tailed paired Student’s t-test. Source data are provided as a Source Data file. e Western blot analysis of the indicated proteins in MIN6 cells. f Fluorescence immunocytochemistry for LC3, in MIN6 cells treated with CPA for 18 h. Nuclei are indicated with staining in blue. Scale bar is 20 μm. g upper, schematic representation of 12 different MIN6 treatments with CPA, MG132 or Baf A1. Green lines indicate CPA treatment. CPA was washed out between 18–24 h; lower, Western blot analysis for proinsulin by non-reducing PAGE electrophoresis in extracts prepared from the 12 MIN6 treatments. Western blot analysis for ubiquitin by reducing PAGE electrophoresis.