Dissecting molecular mechanisms underlying ferroptosis in human umbilical cord mesenchymal stem cells: Role of cystathionine γ-lyase/hydrogen sulfide pathway

Figure 6 Effect of cystathionine γ-lyase/hydrogen sulfide pathway on nuclear factor erythroid 2-related factor 2 activation and ferroptosis in human umbilical cord mesenchymal stem cells. A and B: Exogenous hydrogen sulfide (H₂S) production; C and D: Nuclear factor erythroid 2-related factor 2 (Nrf2) activity; E and F: Western blot analysis and quantification of cystathionine γ-lyase (CSE) protein, Kelch-like ECH-associating protein 1 (Keap1), and cytoplasmic and nuclear Nrf2 protein; G: Change of Keap1 S-sulfhydration. 1,4-Dithio-DL-threitol (DTT, 1 mmol/L, 3 h) served as a negative control; H: Effect of DTT on cytoplasmic and nuclear Nrf2 expression. Histone H3 was used for normalization for nuclear proteins, and glyceraldehyde-3-phosphate dehydrogenase was used for normalization for whole and cytoplasmic protein (n = 3). CSE: Cystathionine γ-lyase; H₂S: Hydrogen sulfide; Nrf2: Nuclear factor erythroid 2-related factor 2; HUCMSCs: Human umbilical cord mesenchymal stem cells; Fer-1: Ferrostatin-1; sh-CSE: Short hairpin RNA targeting cystathionine γ-lyase; Keap1: Kelch-like ECH-associating protein 1; DTT: 1,4-Dithio-DL-threitol; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase. ^bP < 0.01.