Fig. 1: pVHL regulates of mitochondrial mass independent of HIFα.
a, Volcano plot of proteins detected by nanoLC–MS/MS in human PPGL tumours (VHL-mutant/wild type). The dotted line indicates a P value of 0.05 (−log10 P value = 1.3) in two-tailed unpaired t-test. b, Histogram of mitochondrial proteins regulated in human VHL-mutant compared to VHL wild-type PPGL tumours. c, Heat map of the top 50 downregulated and upregulated proteins in human VHL-related PPGL tumours (VHL-mutant/wild type). Red asterisks indicate mitochondrial proteins. d, Top 5 biological processes of the top 50 upregulated (red) and downregulated (green) proteins for human VHL PPGL tumours. e, Immunoblot of 786-O cells expressing pVHL WT or type 2C pVHL mutants (p.Leu188Val or p.Arg64Pro). n = 3 biological independent experiments. f, The corresponding immunofluorescence images are shown. Cells were stained by MitoTracker Red (top). Flow cytometry analysis of MitoTracker Green-stained cells (bottom). Data are presented as mean values ± s.d. One-way analysis of variance (ANOVA) Tukey’s multiple-comparison test. ****P < 0.0001. n = 3 biological independent experiments. g, The volcano plot shows proteins detected by nanoLC–MS/MS in human ccRCC cells (786-O). VHL-null cells (pRC3) were compared to VHL-WT-expressing cells. The dotted line indicates a P value of 0.05 (−log10 P value = 1.3) in a two-tailed unpaired t-test. The histogram shows fold changes of mitochondrial proteins comparing pRC3 to VHL-WT with indicated median value of log2(fold change) −0.27. h, The volcano plot shows proteins detected by nanoLC–MS/MS in 786-O cells. VHL-L188V mutant cells were compared to VHL-WT-expressing cells as in g. The histogram shows fold changes of mitochondrial proteins comparing VHL-L188V to VHL-WT with indicated median value of log2(fold change) of −0.22. i, Immunoblot of 786-O cells stably transfected to produce the indicated pVHL species. j, Immunoblot of VHL MEFs with indicated genotype. In i and j, n = 3 biological independent experiments. k, Corresponding immunofluorescence of VHL MEFs. Cells were stained by MitoTracker Red to visualize mitochondria (top). Flow cytometry analysis of MitoTracker Green-stained MEFs (bottom). n = 3 biological independent experiments. Data are presented as mean values ± s.d. One-way ANOVA Tukey’s multiple-comparison test. ****P < 0.0001.
