Fig. 3. SNX25 in macrophages derived from BM contributed to pain sensation.
a, Confocal microscopy of the plantar skin of the naive hind paw of WT mice, immunolabeled for PGP9.5 and MHC-II. Representative of three independent experiments. Scale bar, 50 μm. b, Replacement rates of myeloid cells by transplanted BM of GFP mice in peripheral blood plotted against time after BMT (n = 4). c, Confocal microscopy of hind paw skin labeled for GFP and MHC-II in WT mice that received BM from GFP mice imaged at weeks 1, 3, 4, 5, 7 and 10 after BMT. Arrowheads denote double-labeled cells. Representative of three independent experiments. Scale bar, 100 μm. d, Confocal microscopy of hind paw skin labeled for GFP (Alexa 594) and Cx3cr1 mRNA (fluorescent in situ hybridization) in WT mice that received BM from GFP mice at week 5 after transplantation. Arrowheads show BM-derived GFP+ cells positive for Cx3cr1 mRNA. Representative of two independent experiments. Scale bar, 100 μm. Bottom, magnified views of the boxed area in the upper panel. Scale bar, 50 μm. e, Flow cytometry strategy to sort donor-derived macrophages (MHC-II+ or F4/80+) using propidium iodide (PI), CD45, F4/80, MHC-II and GFP expression from hind paw skins of WT mice that received BMT from GFP mice. FSC, forward scatter; SSC, side scatter. f, Percentage of GFP+ cells among MHC-II+ or F4/80+ cells. Results are presented as mean ± s.e.m. of three different mice that received BMT from GFP mice. Values are 85.1% and 80.8%, respectively. g, VF thresholds in Snx25+/− → WT BM chimeras (n = 10, P = 0.015) and WT → Snx25+/− BM chimeras (n = 13, P = 0.049) at day 28 after BMT. g, gram. Results are represented as mean ± s.e.m. Statistical significance was calculated using two-tailed Student’s t-test. *P < 0.05.