A23210

Antibodies used in this paper are listed as following: anti‐FEN1 antibody (42 282, Genetex), anti‐γH2AX antibody (ab26350, Abcam), anti‐GAPDH antibody (264 140, Abmart), anti‐BAX antibody (AB026, Beyotime), anti‐BCL‐XL antibody (AB126, Beyotime), anti‐BCL‐2 antibody (AB112, Beyotime), Dy Light 594 Goat‐anti Rabbit (A23420, Abbkine), Dy Light 488 Goat‐anti mouse (A23210, Abbkine).

A group of sections were randomly selected from the three additional groups of sections from each mouse. The sections were rinsed with PBS-T, successively infiltrated with 2 mol/L HCl for 10 min (only required for the anti-BrdU antibody), subjected to antigen retrieval in citrate buffer (0.01 M, 99°C) for 30 min, and incubated with normal goat serum for 2 h at 37°C. Then, the primary anti-CNPase (ab6319, Abcam), anti-Olig2 (rabbit, ab109186, Abcam), anti-SOX10 (ab155279, Abcam), anti-bromodeoxyuridine (BrdU; ab6326, Abcam), anti-5-HT1AR (ab85615, Abcam), anti-platelet-derived growth factor alpha receptor (PDGFαR; ab96569, Abcam), anti-Aβ (ab11132, Abcam), and anti-CDKN2A/p16INK4a (p16; ab201980, Abcam) antibodies were added at a dilution of 1:500 in PBS, incubated at 4°C for 72 h and then rewarmed at 37°C for 2 h. The appropriate DyLight 405, DyLight 488, and DyLight 549-conjugated secondary antibodies (A23140, A23210, and A23320, respectively, Abbkine, P. R. China) were incubated with the sections at a 1:200 dilution. Finally, the sections were mounted on gelatin-coated slides with antifade solution to reduce fluorescence quenching.

Sections of paraffin-embedded tissue were heated at 65°C for 2 h, separated in xylene, and rehydrated in graded ethanol at room temperature. Triton X-100 was applied for 10 min after washing with PBS three times. Sections were washed three times in PBS and then microwaved with a sodium citrate buffer (PH = 6). A wet chamber was used to incubate the slices in 3% hydrogen peroxide for 10 min after three washes in PBS. The tissue sections were washed three times in PBS and treated for 30 min with normal goat serum at 37°C. At 4°C, muscle sections were then incubated overnight with primary antibody (MEF2C, Santa, sc-518152, 1:200). The next day, secondary antibodies were incubated at 37°C for 30 min with dye-labeled secondary antibodies (Abbkine, A23210). After three rounds of washing with PBS, tissue sections were treated with DAPI for 5 minutes and covered.