Fig. 6

Microglial AhR deletion affects the clearance of myelin debris and phagocytic capacity of microglia. A Representative confocal immunofluorescent images of degraded MBP (dMBP) staining in the corpus callosum of AhR^fl/fl mice and CX3CR1-AhR^−/− mice after a 5-week cuprizone administration. Scale bar: 40 μm. B Representative Oil red O stained images in the corpus callosum of AhR^fl/fl mice and CX3CR1-AhR^−/− mice after 5 weeks of cuprizone treatment. Scale bar: 50 μm. C Western blot analysis of AhR protein level in WT primary microglia treated with PBS or purified myelin. 30 μg protein was loaded per well. D The quantification of AhR protein expression in C (n = 4 biological replicates). Data are shown as mean ± SEM and analyzed by unpaired two-tailed t-test. E Representative confocal immunofluorescent images displaying the uptake of pHrodo green bioparticles by WT primary microglia and AhR^−/− primary microglia with or without cytochalasin D treatment. Scale bar: 20 μm. F Flow cytometry analysis of the phagocytosed pHrodo green bioparticles by WT primary microglia and AhR^−/− primary microglia with or without cytochalasin D treatment (n = 3 biological replicates). G The quantification of positive cells engulfing pHrodo green bioparticles and FITC fluorescent intensity in 4 experimental groups (n = 3 biological replicates). Data are shown as mean ± SEM and analyzed by two-way ANOVA with Tukey’s multiple comparisons test. H Flow cytometry analysis of the phagocytosed pHrodo bioparticles by WT primary microglia treated with DMSO or agonist I3S. I The quantification of positive cells engulfing pHrodo green bioparticles and FITC fluorescent intensity in 2 experimental groups (n = 3 biological replicates). Data are shown as mean ± SEM and analyzed by unpaired two-tailed t-test. *p < 0.05, **p < 0.01, ***p < 0.001