Rat sciatic nerves were fixed in situ in 4% PFA for 10 min, dissected, embedded in O.C.T. Compound (Tissue Freezing Medium; Solarbio, Shanghai, China). Sciatic nerve cryosections (5-μm thick) were incubated with acetone for 10 min at − 20 °C, washed in PBS/0.1% Tween 20, blocked for 30 min at room temperature (RT) in blocking buffer (0.3% Triton X-100/10% goat serum/phosphate buffer saline ¼ PBS), and incubated with primary antibodies overnight at 4 °C in blocking buffer. Sections were then washed 3 times in blocking buffer, and sections were incubated with secondary antibodies for 1 h at RT in the dark. Sections were washed again, incubated with DAPI for 5 min at RT, washed and mounted in Citifluor (Agar Scientific).
The primary antibodies used for IF were as follows: neurofilament (1:1000, Abcam, ab8135), SCG10 (1:500, Abcam, ab115513), IBA1 (1:100, Abcam, ab178847), and CD68 (1:100, Abcam, ab125212). All secondary antibodies were also purchased from Abcam. Images were acquired using a Leica TCS SP-II confocal microscope.
The primary antibodies used for IF were as follows: neurofilament (1:1000, Abcam, ab8135), SCG10 (1:500, Abcam, ab115513), IBA1 (1:100, Abcam, ab178847), and CD68 (1:100, Abcam, ab125212). All secondary antibodies were also purchased from Abcam. Images were acquired using a Leica TCS SP-II confocal microscope.