Fig. 6

Assessment of mitochondrial injury induced by GO, MCLR and GO-MCLR in HaCaT and L02 cells. a Mitochondrial ROS were detected by MitoSOX. The HaCaT cells and L02 cells exposed to diverse concentrations of GO, MCLR and GO-MCLR for 24 h were co-incubated with MitoSOX probe for 10 min at a concentration of 5 µM and a dilution ratio of 1:1000. Then, the cells were rinsed 3 times with PBS preheated at 37 ℃. MtROS production levels were observed with Zeiss microscope at excitation wavelength of 510 nm and emission wavelength of 580 nm, and red fluorescence indicated the generation of mtROS. b MMP identified by JC-1 probe. The fluorescent images were obtained by fluorescence microscope at excitation wavelength of 514 nm and emission wavelength of 529 nm for JC-1 monomer, excitation wavelength of 585 nm and emission wavelength of 590 nm for JC-1 aggregates. c Mitochondrial ATP generations in HaCaT and L02 cells treated by 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) were measured by the ATP assay kit using a microplate luminescence detector. d The level of Ca²⁺ were identified in HaCaT/L02 cells stained with Fluo-3AM fluorescence probe. The infected cells were added with 1 mL Fluo-3AM probe diluted in serum-free medium (1:1000) and incubated for 1 h. Intracellular Ca²⁺ concentration was observed at excitation wavelength 506 nm and emission wavelength 526 nm by fluorescence microscopy. *P < 0.05 versus Control, #P < 0.05 versus GO group