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-GSK3β, β-Catenin, and p-ERK were upregulated, whereas FOXO3a and PTEN were downregulated in TRPM2-S-expressing cells compared with control cells

Fig 3: TRPM2-S increased the susceptibility to apoptosis of RPLS cells under oxidative stress. a Representative images of PI staining (upper panels), the analysis indicated that the apoptosis rate was significantly higher in TRPM2-H2O2 cells than control-H2O2 cells and had a concentration-dependence property (down panels). b ROS kit assays showed that ROS was gently increased in TRPM2-S-expressing cells but was sharply elevated in TRPM2-H2O2 cells compared with control cells and control-H2O2 cells respectively. c Western blotting revealed that cleaved caspase 3 was upregulated in TRPM2-S-H2O2 RPLS cells compared with control-H2O2 cells

Fig 4: NAC appeased the TRPM2-S enhanced apoptosis of RPLS cells under oxidative stress. a–e Representative images of flow cytometry for apoptosis, including 94T778-Control (a), 94T778-Control-H2O2 (b), 94T778-TRPM2-S (c), 94T778-TRPM2-S-H2O2 (d), 94T778-TRPM2-S-H2O2-NAC (e). f Analysis of apoptosis rate of 94T778 cell. g–k Representative images of flow cytometry for apoptosis, including SW872-Control (g), SW872-Control-H2O2 (h), SW872-TRPM2-S (i), SW872-TRPM2-S-H2O2 (j), SW872-TRPM2-S-H2O2-NAC (k). l Analysis of apoptosis rate of SW872 cell. m Western blotting revealed that cleaved caspase 3 was upregulated in TRPM2-S-H2O2 RPLS cells but was downregulated in TRPM2-S-H2O2-NAC RPLS cells compared with control-H2O2 and TRPM2-S-H2O2 RPLS cells respectively

Fig 5: Schematic diagram of the molecular mechanism of TRPM2-S in RPLS