Skip to main content
. 2020 May 21;11(5):385. doi: 10.1038/s41419-020-2594-x

Fig. 4. Inhibition of sEH attenuated HG induced mitochondrial dysfunction, ER stress and apoptosis in HK-2 cells.

Fig. 4

a Representative images of MitoTracker Red staining showing the mitochondrial morphology in HK-2 cells after treated with HG or/and t-AUCB for 24 h. b Quantification of percentage of HK-2 cells with fragmented mitochondria (n = 4, *P < 0.05 vs. LG, #P < 0.05 vs. t-AUCB, ΔP < 0.05 vs. HG). c Western blot analysis of Drp1, Mfn2 in HK-2 cells after treated with HG or/and t-AUCB for 24 h. d, e Densitometric analysis of Drp1 and Mfn2 (n = 3, *P < 0.05 vs. LG, #P < 0.05 vs. t-AUCB, ΔP < 0.05 vs. HG). f Representative images of JC-1 staining showing the ΔΨm in different groups of HK-2 cells. g Quantification of the fluorescence intensity of JC-1 staining (n = 3, *P < 0.05 vs. LG, #P < 0.05 vs. t-AUCB, ΔP < 0.05 vs. HG, &P < 0.05 vs. HG + t-AUCB). h, i HK-2 cells were treated with HG or/and t-AUCB for 24 h, and then incubated with DCFH-DA and MitoTracker Red, or Mito SOX and MitoTracker Green. j, k Quantification of the fluorescence intensity of DCFH-DA and Mito SOX staining in figures h and i (n = 3, *P < 0.05 vs. LG, #P < 0.05 vs. t-AUCB, ΔP < 0.05 vs. HG). l Western blot analysis of Bax, Cyt c and cleaved-caspase3 in HK-2 cells after treated with HG or/and t-AUCB for 24 h. m Flow cytometry analysis of the cell apoptotic rate. n–p Densitometric analysis of Bax, Cyt c and cleaved-caspase3 in figure l. q Quantification of the cell apoptotic rate in figure m (n = 3, *P < 0.05 vs. LG, #P < 0.05 vs. t-AUCB, ΔP < 0.05 vs. HG). r Representative images of immunofluorescence double-labeling P62 and Ubiquitin in different groups of HK-2 cells after treated with HG or/and t-AUCB for 24 h. s Colocalization analysis of immunofluorescence images of GFP-LC3 (green) and Ubiquitin (red) in different groups of HK-2 cells after treated with HG or/and t-AUCB for 24 h. t Western blot analysis of Bip, Ire1α, PERK, ATF4, Chop, Caspase12 in HK-2 cells after treated with HG or/and t-AUCB for 24 h. u–z Densitometric analysis of Bip, Ire1α, PERK, ATF4, Chop and Caspase12 in figure t (n = 3, *P < 0.05 vs. LG, #P < 0.05 vs. t-AUCB, ΔP < 0.05 vs. HG).