Fig 2: qRT–PCR and western blot analysis. A–C The top bar graph presents the mRNA/protein levels (Uchl1/UCHL1, Tmprss13/TMPRSS13, and Cst3/CST3) in the diaphragm for the MV and control groups. The mRNA expression levels of three genes measured by quantitative real-time PCR were normalized with respect to ß-actin and are expressed as fold changes relative to the control levels. The protein expression levels were also normalized with respect to ß-actin. The bottom panel presents the western blotting results for UCHL1, TMPRSS13, CST3, and ß-ACTIN protein levels in the diaphragm for the MV and control groups. The data are presented as the mean ± SD (n=6); “*” indicates P < 0.05 compared with the control group

Fig 3: Venn diagram utilized to screen for the co-differentially expressed hub genes. The Venn diagram shows the five overlapping genes (Trim63, Fbxo32, Uchl1, Tmprss13, and Cst3) present among both the differentially expressed genes and the hub genes from the turquoise and yellow modules

Fig 4: Accumulation of CGRP-ir in nerve fibers innervating oral cancer inoculated into the paw of mice. (a) The sciatic nerve was ligated distal to convergence of spinal nerves L4–L6 in mice 3 weeks after inoculation of HSC-3 cancer cells in the paw. The sciatic nerve sample was collected 24 h after ligation (red arrow). (b) Co-localization of CGRP-ir (green) inside the axons (PGP9.5-ir, red) of the sciatic nerve segment. Arrows indicate the colocalization of CGRP-ir (green) inside the axons (red). Scale bar in (b,c) = 10 µm. (c) CGRP-ir in the sciatic axons from sham operated mice (no SNL, n = 3 mice, n = 1829 axons) compared to sciatic nerve ligation mice (SNL, n = 4 mice, n = 2949 axons), t(4776) = 4.88, ** p < 0.01, unpaired Student’s t-test.