Fig. 6: UC-MSC-CM protects L02 hepatocytes and induces mitophagy in L02 cells exposed to H/R conditions.

L02 cells were respectively treated with normal cell culture medium, PBS or UC-MSC-CM and exposed to H/R conditions. a Cell apoptosis was determined through the flow cytometric detection of PI/Annexin V. The data are presented as the means ± SEMs (n = 3/group). b The level of mtROS generation in each experimental group was detected by MitoSOX Red staining and analyzed by flow cytometry. The data are presented as the means ± SEMs (n = 3/group). c The level of ATP production in each group was measured. The data are presented as the means ± SEMs (n = 3/group). d For the assessment of mitochondrial injury, the level of mtDNA in L02 cells in each group was detected. The data are presented as the means ± SEMs (n = 3/group). e The △Ψm of the inner mitochondrial membrane of L02 cells in each group was detected through the flow cytometric assessment of JC-1 staining. The data are presented as the means ± SEMs (n = 3/group). f The coimmunofluorescence of LC3II (fluorescent green) and TOM20 (fluorescent red) was assessed to measure the level of mitophagy in L02 cells in each group. Scale bar: 10μm. g The expression of TOM20 and the LC3II/I ratio in L02 cells in the various groups were determined by western blot assay. The average intensities of the band in the western blots were quantified using GAPDH as an internal reference. The data are presented as the means ± SEMs (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001 (all p-values were obtained by one-way ANOVA).