Journal: Communications Biology

Article Title: Anterior cingulate cortex mediates the comorbidity between colorectal cancer and depression-like behaviors

doi: 10.1038/s42003-025-08719-z

Figure Lengend Snippet: A Representative images of parasympathetic nerve markers VAChT staining in the colorectal tissue. VAChT (Green) and DAPI (blue). B Representative images of sympathetic nerve markers TH staining in the colorectal tissue. TH (Green) and DAPI (blue). Scale bar: 100 μm. C Quantitative of the density of VAChT-labeled parasympathetic nerve in the colorectal tissue across groups (n = 5,8,12,6 per group). D Quantitative of the density of TH-labeled sympathetic nerve in the colorectal tissue across groups (n = 5,10,12,6 per group). E Representative images of sensory nerve markers CGRP staining in the colorectal tissue. CGRP (Green) and DAPI (blue). F Representative images of peptiderergic nerve markers VIP staining in the colorectal tissue. VIP (Green) and DAPI (blue). Scale bar: 100 μm. G Quantitative of the density of CGRP-labeled sensory nerve in the colorectal tissue across groups (n = 4,10,8,6 per group). H Quantitative of the density of VIP-labelled peptiderergic nerve in the colorectal tissue across groups (n = 5,8,11,6 per group). I Representative images of cell proliferation markers Ki67 staining in the colorectal tumor tissue. Ki67 (Green) and DAPI (blue). Scale bar: 100 μm. J Representative images of apoptosis markers Cleaved-caspase3 staining in the colorectal tumor tissue. Cleaved-caspase3 (Green) and DAPI (blue). Scale bar: 100 μm. K Quantitative of the fluorescence intensity of the Ki67 in the colorectal tumor tissue across groups (n = 10,10,6 per group). L Quantitative of the fluorescence intensity of the Cleaved-caspase3 in the colorectal tumor tissue across groups (n = 11,10,5 per group). M Representative Western blot images showing the expression of Ki-67, PCNA, and cleaved caspase-3 in tumor tissues from CRC+Ctrl-ACC (Ctrl), CRC+Gi-ACC (Gi), and CRC+Gq-ACC (Gq) mice. N – P Quantification of Ki-67, PCNA, and cleaved caspase-3 protein levels normalized to β-Tublin in each group, (n = 5,5,6 per group; n = 5,5,6 per group, n = 5,6,6 per group). Q Representative images of tumor size in CRC+Ctrl-ACC, CRC+Gi-ACC and CRC+Gq-ACC groups. R Quantification of tumor volume in CRC+Ctrl-ACC, CRC+Gi-ACC and CRC+Gq-ACC groups, (n = 5,6,6 per group). All data are presented as mean ± s.e.m of the fold change of the Sham+Ctrl-ACC group and analyzed by one-way ANOVA followed by Tukey’s multiple comparisons test ( C , D , G , H ). All data are presented as mean ± s.e.m of the fold change of the CRC+Ctrl-ACC group and analyzed by one-way ANOVA followed by Tukey’s multiple comparisons test ( K , L , N – P , R ). The source data are provided as Supplementary Data in the Figshare repository.

Article Snippet: After sealing at room temperature for 2 h, tip off the blocking solution, drain the excess water on the filter paper pad, place it flat in the wet box, add 200 μL primary antibody (diluted in the blocking buffer) for VAChT (Synaptic Systems, Germany; cat#139103; 1:500), TH (Cell Signaling Technology, USA; cat#E2L6M; 1:500), CGRP (Cell Signaling Technology, USA; cat#14959; 1:500), VIP (Cell Signaling Technology, USA; cat#63269; 1:500), Ki67 (Cell Signaling Technology, USA; cat#9129; 1:500) and cleaved caspase-3 (Cell Signaling Technology, USA; cat# 9664; 1:500) and incubate at 4 °C for 12 h, and then incubate at room temperature for 4 h. On the second day, the colorectal frozen sections were rinsed with 0.4% PBST three times at room temperature for 10 min each time.

Techniques: Staining, Labeling, Fluorescence, Western Blot, Expressing