Fig. 6

Knockout of PINK1 inhibited LPS-induced mitochondrial dynamics and Mdivi-1 reversed the role of PINK1 knockout on DC function. Immature DCs from WT mice were cultured by LPS (1000 ng/mL) and immature DCs from PINK1 knockout mice were cultured by LPS or LPS + Mdivi-1 (20 μM) for 4 h. a–c Representative western blots of Mfn2, OPA1 and COXIV in each group and statistical analysis of relative Mfn2 and OPA1 expression (n = 5). d, e Representative western blots of Drp1 and COXIV in each group and statistical analysis of relative Drp1 expression (n = 5). f, g Representative electron micrographs of mitochondria on DCs in Control, PINK1^−/−, LPS, and LPS + PINK1^−/− group and statistical analysis of average mitochondrial area (n = 3). h, i Representative flow cytometric analysis of MHC-II, CD80, and CD86 expression and statistical analysis of relative MHC-II, CD80, and CD86 expression on DCs in each group (n = 3). j TNF-α mRNA expression was analyzed by PCR and statistical analysis of relative mRNA expression of TNF-α on DCs in each group (n = 3). k IL-12 mRNA expression was analyzed by PCR and statistical analysis of relative mRNA expression of IL-12 on DCs in each group (n = 3). l CD4⁺ T cells were activated with ConA (5 μg/mL) for 18 h, and then were co-cultured with DCs at a ratio of 1:100. CCK-8 was used to test T cell proliferation (n = 5). Results of experiments were shown as the mean ± SD. Statistical significance was assessed using one-way ANOVA analysis with Sidak’s multiple comparisons test (i–l) and two-way ANOVA analysis with Sidak’s multiple comparisons test (b, c, e, f). P values are reported as follows: *, ^#, ^& < 0.05 and **, ^##, ^&& < 0.01