Fig. 4

Microglial activation and RGC loss in response to NMO-IgG injection. a Representative images of retinal cryosections showing microglia (Iba1⁺ green) and iNOS⁺ cells (red) in the HC-IgG and NMO-IgG groups. Top: Merged images showed the retina layers and microglia (Scale bar = 20 µm); Middle: Enlarged image showed the co-localization of Iba1 and iNOS; Bottom: Signal channel of Iba1 and iNOS staining. b Bar graph displaying the number of Iba1⁺ cells per mm² in each group (n = 4 per group, 3 sections per eye). c Bar graph showing the number of Iba1⁺/iNOS⁺ cells per mm² in each group (n = 3 per group, 3 sections per eye). d Bar graph representing the iNOS mRNA levels in the retina on day 1 and day 7 after NMO-IgG injection (n = 3 per group). e Pie chart illustrating the percentage of iNOS⁺ cells among Iba1⁺ cells in the control-IgG and NMO-IgG-treated groups. f Representative image of microglia morphology indicated by Iba1 staining (green) in retinas treated by HC-IgG (top) or NMO-IgG (bottom). g Bar graph displaying the branch length of Iba1⁺ cells in each group (n = 4 per group, 3 sections per eye), (**p < 0.01). h Bar graph displaying the branch number of Iba1⁺ cells in each group (n = 4 per group, 3 sections per eye). i Curve of Iba1⁺ cell Sholl analysis in retinas treated by HC-IgG (green) or NMO-IgG (red). 25 cells per group. j Schematic timeline of PLX3397 treatment, IgG injection, and sample harvesting. k Left: Representative images of Iba1⁺ cells in retinas from normal diet or PLX3397 diet (Scale bar = 1 mm). Right: Higher magnification of the retina (Scale bar = 100 µm). l Visualization of retinal flat mounts showing RBPMS⁺ RGCs in the HC-IgG and NMO-IgG groups on days 7 and 21 with PLX3397 diet. The panoramic view is shown on the top (Scale bar = 1 mm) with a higher magnification of the indicated region displayed on the bottom (Scale bar = 100 µm). m Bar graph illustrating the number of RGCs per mm² in each group (n = 3 per group). **p < 0.01; ***p < 0.001, t-test