Fig. 5: Cx36 inhibits neurotropic virus-induced neuroinflammation, SARM1 activation and behavioral defects in mice.

a Wildtype mice were infused with AAV-PHP.eB virus carrying scramble or Cx36-specific shRNA via tail vain. After eighteen days, the mice were sacrificed, and perfused with 4% PFA for fixation, and brain sections were immunostained for Cx36. Fluorescent images were captured under a Slide Scanner. Scale bar: 1 mm. b Tissues from cortex, cerebellum and brainstem were collected, lysed, and subjected to Western blot analysis to measure Cx36 expression. c Frozen brain sections were immunostained for Iba1, a microglial activation marker. Scale bar: 100 μm. d The number of Iba1-positive cells in c was quantified using ImageJ. e Cortical tissue samples were homogenized with 0.6 M perchloric acid, and the concentration of cADPR was measured using a cycling assay. f The same brain sections were immunostained with anti-CC1 and anti-Tuj1 antibodies to visualize oligodendrocytes and neurons, respectively. Images were captured using a confocal microscope. Scale bar: 50 μm. g The number of CC1-positive cells in f was quantified using ImageJ. h Wildtype and SARM1-KO mice infused with the virus underwent the open field test, during which their track length was recorded over a 30-minute running session. All experiments were repeated at least three times, and data are presented as means ± SDs (n = 3). Statistical significance was determined using Student’s t test in d, e, g and h, following one-way ANOVA for comparisons within groups and two-way ANOVA for comparisons between groups (ns no significance; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001).