Extended Data Fig. 1: Identification of the transgene insertion site in Mlc1Tg mice.
(a) Confocal microscopic images of the dorsal horn of the spinal cord (L4, 30 min after formalin injection, Ipsi: ipsilateral side to injection) immunolabeled for CGRP and c-Fos of WT and Mlc1Tg mice. Arrowheads denoted double-labeled cells. Scale bar, 100 μm. (b) Confocal microscopic images of the dorsal horn of the spinal cord immunolabeled for c-Fos (red) and cell markers (neuron, NeuN; astrocyte, GFAP; microglia, Iba1) of WT mice. Arrowheads denote double-labeled cells. Scale bar, 100 μm. (c) Quantification of c-Fos+ activated neurons in WT and Mlc1Tg mice (WT: n = 3; Mlc1Tg: n = 3, p = 0.005). (d) Diagram showing the insertion site of the RP23-114I6 transgene in chromosome 8 of the Mlc1Tg mice. The positions of three endogenous genes (Snx25, Slc25a4, and Cfap97) relative to the inserted transgene are indicated. (e) mRNA expression levels for Mlc1 (p = 0.856) and Mov10l1 (p = 0.816) in the brain of WT and Mlc1Tg mice (WT: n = 3; Mlc1Tg mice: n = 3). (f) mRNA expression levels for Mlc1 (p = 0.011) and Mov10l1 (p = 0.967) in the spinal cord of WT and Mlc1Tg mice (WT: n = 5; Mlc1Tg mice: n = 4). (g) cDNA microarray data of bone marrow cells of WT and Mlc1Tg mice (WT: n = 3; Mlc1Tg mice: n = 3) were plotted (Y axis: Mlc1Tg; X axis: C57BL/6 WT mice). Snx25, Slc25a4, and Cfap97 are indicated by colored dots. (h) RT-qPCR analyses of Snx25 mRNA in the spinal cord (WT: n = 5; Mlc1Tg mice: n = 4), DRG (WT: n = 5; Mlc1Tg mice: n = 5), and hind paw skin (WT: n = 3; Mlc1Tg mice: n = 5). (i) Immunoblot analysis showing the expression of SNX25 protein in the lung of WT and Mlc1Tg mice. Results are represented as mean ± SEM. Statistical analyses were performed using the two-tailed Student’s t-test. *p < 0.05, **p < 0.01. n.s., not significant. n.d., not detected. Representative of two independent experiments (a and b).
