Fig 2: TRPM2-S promotes proliferation via activating ERK and AKT. a Western blotting indicated TRPM2-S expression in pcDNA3.1-TRPM2-S-Flag cells was obviously higher than in empty vector RPLS cell lines. b TRPM2-S inhibited the intracellular flow of Ca2+. c Growth curves of live-cell imaging showed TRPM2-S-expressing 94T778 and SW872 cells have a higher proliferative capacity than negative control cells. d Representative images of EdU assays (left panels). EdU showed that a significant proliferation rate was observed in TRPM2-S-expressing RPLS cell lines (right panels). e Western blotting revealed that p-Akt, p-GSK3ß, ß-Catenin, and p-ERK were upregulated, whereas FOXO3a and PTEN were downregulated in TRPM2-S-expressing cells compared with control cells

Fig 3: The influence of PTEN on cell damage and apoptosis of HK-2 cells in response to I/R treatment.A Western blotting analysis showing changes in PTEN and NGAL in I/R-treated HK-2 cells under an antimycin A concentration gradient. B Quantification of cell viability (%) by the CCK-8 assay in I/R-treated HK-2 cells under an antimycin A concentration gradient. C Western blotting analysis showing changes in PTEN and NGAL in I/R-treated HK-2 cells at the indicated reperfusion times. D Quantification of cell viability (%) by the CCK-8 assay in I/R-treated HK-2 cells at the indicated reperfusion times (**P < 0.01 and ***P < 0.001 compared with values in the Ctrl group). E Western blotting analysis showing changes in PTEN and NGAL in I/R-treated HK-2 cells in the absence and presence of si-PTEN. F Quantification of cell viability (%) in I/R-treated HK-2 cells in the absence and presence of si-PTEN. G Western blotting analysis showing changes in PTEN and NGAL in I/R-treated HK-2 cells in the absence and presence of LV-PTEN. H Quantification of cell viability (%) in I/R-treated HK-2 cells in the absence and presence of LV-PTEN. I Quantification of cell apoptosis (%) in I/R-treated HK-2 cells in the absence and presence of si-PTEN or LV-PTEN. A white rectangle represents the Q3 quadrant, and a gray rectangle represents the Q2 quadrant.

Fig 4: The effect of renal proximal tubular cell (RPTC)-specific PTEN overexpression on I/R-induced renal injury and apoptosis.A Schematic representation of renal proximal tubular cell (RPTC)-specific PTEN overexpression mice established by crossing PTENfl-stop-fl mice with Ggt1-Cre mice. B Identification of the genotype PTENfl-stop-fl, Cre+/+ by Northern blotting. C Representative immunohistochemical staining of PTEN in kidney in PTENfl-stop-fl, Cre-/- and PTENfl-stop-fl, Cre+/+ mice (scale bar: upper, 200 µm; lower, 50 µm). D Western blotting analysis showing changes in PTEN in different organs in PTENfl-stop-fl, Cre-/- and PTENfl-stop-fl, Cre+/+ mice (n = 8 mice in each group). E Concentration of serum creatinine in PTENfl-stop-fl, Cre-/- and PTENfl-stop-fl, Cre+/+ mice subjected to bilateral renal artery obstruction for 25 min, followed by 24 h of reperfusion (BIRI). F Representative HE staining and TUNEL staining of kidney in PTENfl-stop-fl, Cre-/- and PTENfl-stop-fl, Cre+/+ mice subjected to BIRI (scale bar: upper, 50 µm; lower, 20 µm). G Quantification of the number of TUNEL-positive cells (n = 6 mice in each group). H Western blotting analysis showing changes in PTEN and NGAL in renal cortex in PTENfl-stop-fl, Cre-/- and PTENfl-stop-fl, Cre+/+ mice subjected to BIRI (n = 6 mice in each group).

Fig 5: Mass spectrometry-based proteomics in I/R-induced RPTC-specific PTEN overexpression mice.A The heat map of renal distinct protein expression comparing sham, BIRI, and BIRI + PTEN-OE mice (n = 3 mice in each group). B, C The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment of differentially expressed proteins (DEPs) comparing BIRI versus sham and BIRI + PTEN-OE versus BIRI groups, respectively. D, E The KEGG pathway enrichment combined with protein–protein interaction (PPI) analysis of DEPs, comparing BIRI versus sham and BIRI + PTEN-OE versus BIRI groups, respectively. F Western blotting analysis showing changes in LC3 and P62 in I/R-treated HK-2 cells in the absence and presence of Baf A1. G The heat map of DEPs enriched in the phagosome, endocytosis, autophagy, and lysosome pathways among sham, BIRI, and BIRI + PTEN-OE mice.