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[Preprint]. 2025 May 18:2025.05.14.654046. [Version 1] doi: 10.1101/2025.05.14.654046

Fig. 3: Netrin-1 promotes the innervation of precancerous PanIN lesions.

Fig. 3:

a-c, Representative immunohistochemistry staining for Netrin-1 and Sox9 in pancreatic sections through PanIN (b), PDAC tumor (c) and adjacent asymptomatic tissue (a) obtained from 6.5-week-old KIC mice. DAPI nuclear staining. Scale bar: 50 μm.

d, Quantification of Netrin-1 fluorescence labeling in PanIN (P), PDAC tumor (T), and adjacent tissue (A). Data are presented as mean ± SEM. n = 15 PanINs, 16 PDAC, and 16 asymptomatic ROIs from 3 KIC mice. p-values are indicated on the graph (Kruskal–Wallis test).

e-f, Representative RNAscope in situ hybridization for Th mRNA and Dcc mRNA in sections of the celiac superior mesenteric ganglia (CSMG) from 6.5-week-old control (e) and KIC (f) mice. Higher magnification insets highlight individual sympathetic neuronal cell bodies. Scale bars: 20 μm and 5 μm (insets).

g, Quantification of Dcc mRNA (dots) in CSMG sympathetic neurons from 6.5-week-old control (ctr) and KIC mice. Data are presented as mean ± SEM. n =5 CSMG sections from 3 control mice and 6 CSMG sections from 3 KIC mice. P-values are indicated on the graph (Mann–Whitney test).

h, Experimental design for the treatment of KIC mice with either anti-Netrin-1 antibody (αNetrin-1) or isotype control antibody (Iso-antibody).

i-l, left panels in (i) and (k) show maximum intensity projection of 3D images of TH+ sympathetic networks in PanIN and adjacent tissue treated with Iso-antibody (i) or αNetrin-1 (k) antibodies. Right panels in (i) and (k) show 2D optical sections of PanINs identified by tissue autofluorescence (AF). (j) and (l) show the 3D reconstruction of sympathetic axons and branching points (purple dots) in PanINs and adjacent tissues. The volume of the PanIN is shown in yellow. Scale bars: 100 μm (i, k, and left panels of j and l), 20 μm (right panels of j and l).

m-n, 3D quantification of TH+ sympathetic axon density (m) and branching (n) in PanIN (P) and adjacent (A) tissues of KIC mice treated with Iso-antibody (Iso) or αNetrin-1 (αNet). Data are presented as mean ± SEM. n = 6 PanINs and 6 asymptomatic ROIs from 3 mice per group. Significant p-values are indicated on the graphs (ns: p > 0.05) (Kruskal–Wallis test).