Fig. 4: OTUD5 protects renal tubular cells from ferroptosis in response to H/R injury.
From: Autophagy of OTUD5 destabilizes GPX4 to confer ferroptosis-dependent kidney injury
a Ferroptosis was measured using fluorescent lipid peroxidation sensor BODIPY™ 581/591 C11 and cell death Dye 7AAD in siControl or siOTUD5-transfected cells 3 h after H/R induction (n = 5 independent experiments). b Ferroptosis was measured in empty vector (EV), WT OTUD5, or enzymatically inactive OTUD5 (C224S) plasmid-transfected HK2 cells after H/R induction (n = 5 independent experiments). c Cell viability was measured in siControl or siOTUD5-transfected cells at 24, 48, and 72 h after ex vivo I/R induction (n = 5 independent experiments). d Cell viability was measured in EV, WT OTUD5, or OTUD5-C224S-transfected cells after H/R induction (n = 5 independent experiments). e Representative images and quantification of PRTCs ferroptosis from WT and Pax8CreOtud5fl/fl mouse stained by liperfluo; white arrowheads represent the broken nucleus in ferroptotic cells (n = 5 independent experiments), scale bars: 50 μm. The arrowhead indicates the broken nucleus. f Cell ferroptosis was measured in WT OTUD5-transfected cells, either in the presence or absence of various doses of the GPX4 inhibitor RSL3, under H/R induction (n = 5 independent experiments). g GSH/GSSG ratio and intracellular GSH level were measured in siControl or siOTUD5-transfected cells after H/R induction (n = 3 independent experiments). h GSH/GSSG ratio and intracellular GSH level were measured in EV or WT OTUD5 plasmid-transfected cells after ex vivo I/R induction (n = 3 independent experiments). Data are presented as mean ± s.e.m.; all statistical significance between groups as indicated was determined using an unpaired two-tailed Student’s t-test.
