Fig. 5

Knockout of PINK1 inhibited LPS-induced mitophagy in DCs and Parkin reversed the role of PINK1 knockout on DC function. Immature DCs from WT mice and PINK1 knockout mice were cultured by PBS or LPS (1000 ng/mL) for 4 h. a, b Representative western blots of PARK2 and COXIV in each group and statistical analysis of relative PARK2 expression (n = 5). c–e Representative western blots of LC3, Tomm20 and β-actin in each group and statistical analysis of relative LC3II and Tomm20 expression (n = 5). f, g Representative fluorescence images of the co-localization of lysosome and mitophagy and statistical analysis of percentage of mitophagy-positive cells on DCs in each group, including lysosome (green), mitophagy (yellow) and nucleus (blue) (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 Representative expression of T cell proliferation in each group (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, g). P values are reported as follows: ^& < 0.05 and **, ^##, ^&& < 0.01