Fig. 4: Oncogenic truncated APC drives dysregulation of free cholesterol homeostasis in vivo. | Nature Communications

Fig. 4: Oncogenic truncated APC drives dysregulation of free cholesterol homeostasis in vivo.

From: Mutant APC reshapes Wnt signaling plasma membrane nanodomains by altering cholesterol levels via oncogenic β-catenin

Fig. 4

A CRC mouse model expressing oncogenic APC. B Mouse experimental design. C Representative images of crypts stained with filipin III from AfGC homo mice. Scale bars: 100 µm. D Colon tissue from GC and AfGC homo mice exhibiting polyposis. Arrowhead, mesenteric adipose tissue; arrow, polyp. Scale bars: 1 cm. E H&E staining of colonic swiss roll (top) and quantitative analysis of crypt length. Scale bars: 1 mm; zoom, 125 µm. F Whole colon quantitative fluorescence intensity analysis of filipin III in GC, AGC het, and AfGC homo. G Crypt-derived GPMV quantitative analysis of plasma membrane free cholesterol levels. Representative filipin III fluorescence images of crypt from AfGC (top left), formation of a GPMV (blebbing, white arrow) from a crypt (bottom left), mixed GPMVs (white arrow), and whole cells (green arrow) (top right), and quantitative analysis of GPMV filipin III fluorescence intensity (bottom). Scale bars: 10 µm. H CSC quantitative analysis of filipin III in AfGC homo mice. Representative images of filipin III-stained CSCs (top) from AfGC homo and filipin III quantitative analysis (bottom). Scale bars: 20 µm. I Representation of colon tumor biopsy harvesting a patient’s tumor to generate CRC-PDOs. J PDO quantitative analysis of plasma membrane free cholesterol. Representative images of PDOs and filipin III quantitative intensity analysis. Scale bars: 50 µm. For all experiments, error bars represent E, F crypts from n = 3–6 mice, E (8♂,7♀), F (15♂,15♀), G crypt-derived GPMVs from n = 3–15 mice (15♂,15♀), H CSCs from n = 5–11 mice (13♂,13♀), normalized to WT APC mice or J organoids from n = 21–37 fields of view (FOV) (2–3 PDOs per FOV) (mean ± SD, exact n value is shown in each graph). The exact total number of crypts, GPMVs, CSCs, and FOVs analyzed are provided below each bar. For all experiments, statistical significance was determined by one-way ANOVA and post Tukey’s multiple comparison test. Different letters indicate significant differences between WT APC (control) and mutant APC groups (experimental) at each time point (P < 0.05). Source data are provided as a Source data file.

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