Fig. 4

Altered morphology in the mouse cerebellum infected with lentivirus for Mettl3 knockdown. a, b GFP fluorescence image (a) and hematoxylin-eosin (HE) staining (b) of the mouse cerebellum dissected 10-days post-lentivirus infection. Arrows indicate the position of injection. Scale bar in a represents 1 mm. Scale bar in b represents 200 μm. c Western blot analysis to confirm the knockdown efficiency of Mettl3 in the neuro2a cell line. β-ACTIN was used as an internal control. sh-1 and sh-2 represent two kinds of Mettl3 knockdown lentiviral vectors. sh-1 was used for subsequent in vivo analysis. d Decreased m⁶A levels resulting from Mettl3 knocking down as analyzed using UHPLC-MS/MS. n = 4. *P < 0.05. e, f Immunofluorescence staining with antibodies against Calbindin-D28K (e) and GFAP (f) was performed to detect Purkinje cells (e) and astrocytes (f) upon lentivirus infection (n = 3 biological replicates). GFP fluorescence indicates the area with Mettl3 knockdown. Sections were counterstained with DAPI to visualize the nuclei. Scale bar represents 100 μm