Figure 7

Reconstitution of Nrf2 fully restores GSH and GTP and partially restores cell viability, ATP, NADH, and NADPH levels in TRPM2 depleted cells. SH-SY5Y cells in which TRPM2 was depleted (KO-1 and KO-2) were stably transfected with Nrf2 (KO-Nrf2) or empty vector (KO-v), and scrambled controls cells with empty vector (Scr-v). (a) Cells were treated with media or 0.3 µM doxorubicin for 24 or 48 hours. Cell viability was measured with XTT, and ATP, GSH, NAD⁺, NADH, NADP⁺, and NADPH were quantitated with biochemical assays. Representative results from three experiments are shown. *p < 0.0056, significance determined with the Bonferroni Correction, two-way ANOVA. (b) Metabolomic analysis of TRPM2 depleted cells reconstituted with Nrf2 and treated with doxorubicin for 24 hours was performed (n = 6 replicates/group). Levels of ATP were partially restored, and GTP and GSH fully restored by expression of Nrf2. *p < 0.05, ** < 0.01, *** < 0.001, **** < 0.0001, one-way ANOVA.