Fig. 5
TMEM106B drives nuclear translocation of TFEB and regulates TFEB-mediated expression of lysosomal genes. a 393P cells with inducible expression of TMEM106B or GFP alone were transfected with Mcherry-TFEB construct. After induction for 24–48 h, cells were counter stained with Hoechst33342. Live cells were imaged in real time. Indicated scale bar = 5 µm. b Western blot analysis upon subcellular fractionation of cells with induction of GFP-TMEM106B or GFP alone as control. c, d qPCR analysis for expression of lysosomal genes upon expression or knockdown of TMEM106B expression. e Heatmap of intergene expression correlation across 1016 lung cancers from TCGA49. Color bar represents Pearson’s r value (based on log-transformed values); red is positive correlation. f Western blot analysis after transfection of either control siRNA or siRNA against TFEB in cells expressing either GFP alone or GFP-TMEM106B. g qPCR analysis for expression of lysosomal genes upon induction of GFP alone as control (Vec) or TMEM106B expression, after transfection of either control siRNA or siRNA against TFEB in cells expressing either GFP or GFP-TMEM106B. h Relative cathepsin B or cathepsin K activity as measured by quantification of fluorescence intensity after staining with Magic Red Cathepsin B or K reagent to assess for activity of cathepsin B or K, in cells as described in g. All asterisks indicate statistical significance by t test (n ≥ 3, *p ≤ 0.05)
