Figure 3

Western blot and immunofluorescence confirmation of Il6ra^−/− in isolated Müller glial cells (MGCs). (A) Knockout of Il6ra was assessed by western blot of MGCs isolated from wildtype (WT), floxed littermates (fl/fl), and MGC-specific Il6ra^−/− (KO) mice. Purified IL-6Rα protein (70 kDa) was used as a positive control (+ ctrl) for the membrane-bound receptor. Western blots were also probed for β-actin (42 kDa) for normalization. IL-6Rα protein expression was observed in WT (n = 3) and floxed (n = 1) MGC lysates but absent in KO MGCs (n = 5, FC = 0.156). Data is represented as fold control mean ± SD, n = 3–5/group, ****p-value < 0.0001 vs. WT (unpaired T-test). (B) Immunofluorescence staining for IL-6Rα (red) and vimentin (green) in isolated MGCs from WT and KO mice, counterstained with DAPI (blue). Images were captured using a Leica STELLARIS Confocal Microscope at 40× magnification (n = 4 technical replicates/group, n = 5–10 images/well).