Rat anti ki 67

Rat anti-CD31 (BD Biosciences, #553370) at 1:200 and Rat anti-CD31 (Dianova, #DIA-310) at 1:100, mouse anti-SMA-α (Chemicon, #CBL171) at 1:200, rat anti-Ki-67 (Abcam, #ab15580) at 1:500, goat anti-chemerin (R&D Systems, #AF2325) at 1:100, mouse anti-MAB-1 (Hypoxyprobe-1, #Mouse-Mab1) at 1:100, rabbit anti-PPAR-γ (Abcam, ab#59256) at 1:500, rabbit anti-β-actin (Santa Cruz Biotechnology, #sc-47778) at 1:1,000 and rabbit anti-NKI-A59 (a gift from B. Floot, Netherlands Cancer Institute, Amsterdam, Netherlands) at 1:100 were used as primary antibodies.

For immunofluorescence staining, the following primary antibodies were used: rabbit anti‐calcium binding adaptor molecule 1 (IBA1) (Wako), goat anti‐GFAP and anti‐NeuN (Abcam), mouse anti‐IL‐6, rabbit anti‐TNF‐α (Santa Cruz), rat anti‐KI67 (Abcam), mouse anti‐FOXP3 (BD), mouse anti‐CD16, and anti‐CD206 (Boster). The slices were imaged using a confocal microscope (Olympus FV1000). Three‐dimensional reconstruction and co‐localization analysis were conducted using Imaris software. Morphological changes in microglia were analyzed with the Sholl analysis plugin for ImageJ. Detailed information about three‐dimensional reconstruction and measurement of the co‐localization coefficient using Imaris software, and the Sholl analysis is thoroughly demonstrated in Appendix S1.

Murine skin tissue samples were embedded in paraffin, sectioned, and then stained with H&E or Masson’s trichome at different time points, according to routine procedures. For immunofluorescence analyses, skin tissue sections were incubated with the following primary antibodies: rat anti-mouse α-smooth muscle actin (α-SMA; 1:100; Abcam, Cambridge, United Kingdom), rat anti-mouse p75 (1:50, Abcam), rabbit anti-mouse MBP (1:1000, Abcam), rabbit anti-mouse SOX10 (1:100, Abcam), rabbit anti-mouse p75 (1:100, Abcam), rat anti-Ki67 (1:100, Abcam), and rabbit anti-mouse c-Jun (1:100, Abcam). Images were captured using a fluorescence microscope (IX71FL, Olympus), and the percentage of p75^high MBP^low NBs relative to total NBs, the percentage of p75⁺/SOX10⁺cells relative to total SOX10⁺cells were analyzed. And the numbers of p75⁺/Ki67⁺ cells and c-Jun+/Ki67⁺ cells in different groups were counted. The folded areas in all of these immunohistochemistry images have been excluded from quantification in this manuscript. And non-specific p75 stainings in the image’s folded tissue were also excluded from quantification in this manuscript.