Extended Data Fig. 3: UCP2 has key role in redox homeostasis of PDAC cells.
a, GSH or NAC in the medium fully rescues the growth defect of UCP2-silenced Panc1 cells. b, UCP2 silencing reduces the GSH/GSSG ratio only in KRASmut cells. c–i, UCP2 silencing increases ROS levels only in KRASmut cells. The cytofluorimetric raw data of ROS analysis in control and UCP-silenced cells are shown (d–i). j, UCP2 silencing reduces the NADPH/NADP+ ratio only in KRASMUT cells. The histogram data were referred to CtrshRNA-transfected cells (b, c and j). k, l, In PDAC, UCP2 expression levels are associated to those of genes involved in the redox homeostasis. The GSEA panels show the enrichment of gene sets related to UCP2 expression in roughly 300 PDAC primary tumour samples from E-MTAB-6134 dataset (k) and 43 PDAC cell lines from dataset GSE36133 (l). NES, normalized enrichment score. m, Fluorescence microscopy images of mitochondrial-targeted GFP transfected Patu8988T and BxPC3 cells before and after 4 minutes of incubation with digitonin fractionation buffer. Scale bar = 10 µm. Values represent means ± SD of two biologically independent triplicates. Statistical significance was calculated by unpaired two-tailed Student’s t-test; Welch correction was applied to shRNA1 vs shRNA1/GSH (a), Ctr vs shRNA1 and Ctr vs shRNA2 (Panc1) (c).
