Fig. 4

Ferroptosis caused by GO, MCLR and GO-MCLR in HaCaT and L02 cells. HaCaT and L02 cells were co-incubated with GO (5 µg/mL and 50 µg/mL), MCLR (0.06 µg/mL and 0.6 µg/mL) and GO-MCLR complex (5 µg/mL + 0.06 µg/mL, 50 µg/mL + 0.6 µg/mL) suspensions for 24 h. a After exposure, HaCaT/L02 cells were incubated with the FerroOrange ferrous ion fluorescence probe for 1 h keeping out light. The content of Fe²⁺ in the cells were imaged using the g-excitation filter in fluorescence microscopy. The stronger the fluorescence, the more Fe²⁺ accumulated in the cells. b The level of SLC7A11 and GPX4 expression were analyzed by Western blot. M0.06: MCLR 0.06 µg/mL; G5: GO 5 µg/mL; GM5: GO-MCLR 5 µg/mL + 0.06 µg/mL; M0.6: MCLR 0.6 µg/mL; G50: GO 50 µg/mL; GM50: GO-MCLR 50 µg/mL + 0.6 µg/mL. c Band intensities were quantified and normalized to Tublin. *P < 0.05 versus Control, #P < 0.05 versus GO group