IF
The L3-L5 segments of the spinal cord tissues of rats were fixed for 6 h in 4% paraformaldehyde and then embedded in paraffin. The spinal cord cross-section was cut using a paraffin microtome (3 µm). Primary antibodies used were anti-transmembrane protein 119 (Tmem119, 1:100, Novus Biologicals, New York, USA). The primary antibodies were incubated at 4 ℃ overnight, and subsequently, secondary antibodies were incubated at room temperature for 1 h. Nucleus were then stained by mounting medium with DAPI fluorescent sealing reagent (Elabscience, Wuhan, China) for 3-5 min. The images were observed with a digital pathology image scanner (DM5000B, Leica, Germany).