To further identify the MEF2D, GFAP and caspase‐3 expression, we used immunofluorescence. Paraffin sections were dewaxed, and we repaired antigen and removed endogenous peroxidase. 0.2% Triton X‐100 and 5% bovine serum albumin (BSA) were used to block for 1 h, and sections were incubated at 4°C overnight with anti‐MEF2D (1:200, Abcam, Cambridge, UK, ab246884), anti‐GFAP (1:200, Abcam, Cambridge, UK, ab7260) and anti‐caspase‐3 (1:200, Abcam, Cambridge, UK, ab13847). Then, the sections were incubated with secondary antibody (FITC; 1:100, Abcam, Cambridge, UK, ab150077) for 1 h and stained with 4',6‐diamidino‐2‐phenylindole (DAPI) or propidium iodide (PI) for 6 min in the dark. Finally, the images were captured immediately by the confocal laser scanning microscope (Zeiss, Jena, Germany). Mean fluorescence intensity was used to evaluate the level of each protein expression.

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