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. 2023 Dec 18;14:8393. doi: 10.1038/s41467-023-44228-5

Fig. 6. H/R reduces OTUD5 through repressing mTORC1 signaling.

Fig. 6

a Spatial feature plot and Violin plot of mTOR activity signature score in ST spots of the two groups. b PRTCs were treated by H/R for 3, 6, or 12 h and then collected and analyzed by immunoblotting. c Cells were treated by H/R in the presence of mTOR activator MHY1485 or mTOR inhibitor rapamycin for 3 h and then collected and analyzed by immunoblotting. d HK2 cells were treated with hypoxia condition for 6 h and then collected and analyzed by immunoblotting. e HK2 cells were transfected with siControl or siTSC1 for 48 h and treated with or without hypoxia condition. Cells were collected and analyzed by immunoblotting. f, g HK2 cells were treated with hypoxia conditions in the presence or absence of mTOR activator MHY1485. And cells were collected for ferroptosis detection using flow cytometry. And cells were collected for detecting OTUD5’s autophagy (f, n = 10 independent cells) using IF staining, and cell ferroptosis (g, n = 5 independent experiments) using flow cytometry. The magnification images show the degrading OTUD5 in the lysosome (LC3). Scale bar, 20 μm.; Data are presented as mean ± s.e.m., statistical significance between groups, as indicated, was determined using an unpaired two-tailed Student’s t-test.