Ab245134

For immunohistofluorescence staining, rats were anesthetized and transcardially perfused with 200 mL 0.9% saline, followed by 400 mL 4% paraformaldehyde at seven days post-transplantation (Supplementary Fig. 3a). Thereafter, the spinal cord was dissected (1.0 cm above and below the injured site, Supplementary Fig. 3b) and cryoprotected in 30% sucrose in 0.1 M PBS dehydrate at 4 °C. The tissues were sliced at 10 mm thickness by cryostat and fixed on the PLL-coated slides. The details of immunohistofluorescence staining were consistent with the above-mentioned description (Fan et al. 2019 (link)). The primary antibodies included anti-p75 (ab245134, 1:200, Abcam), GFP (ab6673,1:200, Abcam), Iba1 (019-19741, 1:500, Wako), iNOS (ab49999, 1:200, Abcam), Arg-1 (ab60176, 1:200, Abcam), NF68 (2835s, 1:50, Cell Signaling Technology) and DAPI (ab228549, 1:1000, Abcam). Other following procedures were same as the immunocytofluorescence.

OECs and microglia grown on the coverslips were treated according to the experimental protocols. Then the immunofluorescence was performed following previously described methods (Wang et al. 2022 (link); Guo et al. 2020 (link)). The primary antibodies were used to stain cells, including anti-p75 (ab245134, 1:300, Abcam), Iba1 (019-19741, 1:500, Wako), iNOS (ab49999, 1:300, Abcam), Arg-1 (ab60176, 1:300, Abcam), TREM2 (ab95470, 1:500, Abcam), F4/80(ab6640, 1:200, Abcam), pNF-κB (3033, 1:200, Cell Signaling Technology), APOE (ab183597, 1:200, Abcam) and DAPI (ab228549, 1:1000, Abcam). After incubation with the corresponding secondary antibodies, coverslips were washed and mounted on the slides. All images were captured by fluorescence microscope (Leica Microsystems, Germany).