Fig. 5: Cholinergic niche sustains compensatory PI3K signaling following epithelial cholinergic receptor ablation.
From: Prox1-positive cells monitor and sustain the murine intestinal epithelial cholinergic niche
a EGFR, Erbb2, and Erbb3 expression in intestinal single cells13; TPM transcripts per kilobase million. b Immunoblot analyses of epithelial-enriched Vil-Cre × M3R fl/fl mouse samples for p-EGFR/EGFR, DCLK1, and M3R (n = 3 WT mice p-EGFR/EGFR; 4 WT mice DCLK1, M3R; n = 4 Vil-Cre × M3R fl/fl −/− p-EGFR/EGFR, M3R; 3 Vil-Cre × M3R fl/fl −/− mice DCLK1; WT p-EGFR/EGFR Mean = 0.783, SEM = 0.073; DCLK1 Mean = 1, SEM = 0.234; M3R Mean = 1, SEM = 0.04; Vil-Cre × M3R fl/fl p-EGFR/EGFR Mean = 0.45, SEM = 0.043; DCLK1 Mean = 4.827, SEM = 0.95; M3R Mean = 0.612, SEM = 0.036; multiple t tests; p-EGFR/EGFR t = 4.175, df = 5; DCLK1 t = 4.547, df = 5; M3R t = 7.177, df = 6). c Analysis of Vil-Cre × M3R fl/fl × M1R fl/fl mice shows additive downregulation of EGFR activation compared with Vil-Cre × M3R fl/fl mice (n = 3 Vil-Cre × M3R fl/fl −/− mice, n = 3 Vil-Cre × M3R fl/fl −/− × M1R fl/fl −/− mice; Vil-Cre × M3R fl/fl −/− p-EGFR/EGFR Mean = 1, SEM = 0.076; Vil-Cre × M3R fl/fl −/− × M1R fl/fl −/− p-EGFR/EGFR Mean = 0.260, SEM = 0.080; unpaired t test, two-tailed, t = 6.721, df = 4). d Epithelial M3R ablation sustained PI3K-PDK1 signaling (n = 3 WT, 3 Vil-Cre × M3R fl/fl −/− mice PI3K, PDK1; n = 3 WT mice p-Akt/Akt, n = 4 Vil-Cre × M3R fl/fl −/− mice p-Akt/Akt; WT PI3K Mean = 1, SEM = 0.144; PDK1 Mean = 1, SEM = 0.799; p-Akt/Akt Mean = 1, SEM = 0.103; Vil-Cre × M3R fl/fl −/− PI3K Mean = 1.844, SEM = 0.069; PDK1 Mean = 5.906, SEM = 1.697; p-Akt/Akt Mean = 0.941, SEM = 0.142; multiple t tests; PI3K t = 5.278, df = 4; PDK1 t = 2.617, df = 4, p = 0.059; p-Akt/Akt t = 0.3102, df = 5). Source data are provided as a Source Data file. *p < 0,05, **p < 0.01, ***p < 0,005.
