Extended Data Fig. 1: Tumor innervation predicts poor CRC survival.

(a) Representative images of nerve fibers in human CRC tumors labeled with NF-L/PGP9.5 from 3 patients. (b-e) NF-L⁺ nerve fibers in 70 CRC patients. (b) Representative images of NF-L⁺ nerve fibers in tumor sections. (c) Negative correlation with overall survival. (d, e) Higher density/count of NF-L⁺ nerve fibers predict poorer survival. (f-i) Representative images of NF-L⁺ nerve fibers in xenograft models. (f) CMT93 rectal CDX (male, C57BL/6 J, n = 16 mice). (g) Sciatic nerve CDX (male, C57BL/6 J, n = 3 mice). (h) SW480 rectal CDX (male, nude, n = 12 mice). (i) PDX (male, B-NDG, n = 3 mice): No NF-L⁺ nerve fibers. (j) Comparable Ngf/Gabbr1 mRNA in CMT93 cells (n = 3 biological replicates). (k) GW441756 treatment in CMT93 rectal CDX model. NF-L⁺ nerve fibers in rectal tumors and tumor volome at days 6/10/14 post-injection of cells. At day 6, n = 4 mice per group. At day 10, n = 5 mice per group. At day 14, n = 4 mice per group. The number of NF-L⁺ nerve fibers is the normalized number in the tumor area of 1 ×10⁸ μm². The red arrowheads indicate nerve fibers. Data are presented as mean ± SEM. Two-tailed Pearson Correlation Analysis (c), two-tailed Gehan-Breslow-Wilcoxon test (d, e), two-tailed Mann Whitney test (j, nerve density at days 10/14 in k), two-tailed unpaired student’s t test (nerve density at day 6 in k, tumor volume at days 6/10 in k), and two-tailed unpaired student’s t test with Welch’s correction (tumor volume at day 14 in k) are used. n.s., not significant.

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