s0791  (Selleck Chemicals)

Journal: bioRxiv

Article Title: Slap restricts oncogenic Src-family kinase signaling to maintain colonic epithelial homeostasis

doi: 10.64898/2026.01.05.697659

Figure Lengend Snippet: (A) Slap-dependent colonic organoid development. Representative images of colon-derived organoids from Slap f/f and Slap f/f Villin-Cre ERT2 mice cultured in Matrigel for 2 days. Quantification of organoid number and area is shown as mean ± SEM from 100-200 organoids per mouse (n = 8 mice per group). ****p<0.0001 Mann–Whitney test. (B) Intestinal Slap deletion increases SFK activity and global protein tyrosine phosphorylation in colonic crypts. Representative immunoblots (right) and quantification (left) of phospho-SFK (pSRC) and total phospho-tyrosine levels in lysates from isolated colonic crypts of the indicated genotypes. Data are presented as mean ± SEM from n = 4-5 mice per group. *p<0.05; **p<0.001 t test. (C) Slap-dependent colonic organoid expansion requires SFK activity. Representative images and quantification of colon-derived organoids from Slap f/f and Slap f/f Villin-Cre ERT2 mice cultured for 2 days in the presence or absence of the SRC-family kinase inhibitor eCF506 (100 nM). Data are shown as mean ± SEM from 100-200 organoids per mouse, n = 4 mice per group. ***p<0.001, ****p<0.0001 Mann–Whitney test.

Article Snippet: After Matrigel polymerization, 500 μL of M2 medium (M1 media supplemented with EGF (50 ng/mL, Bio-techne), Noggin (100 ng/mL, Stem cell technologies), R-spondin1 (500 ng/mL, Stem cell technologies), Y27 (10 μM, Sigma-Aldrich), CHIR-99021 (3 μM, Tebu-Bio) and WNT3a (50 n/ mL, Thermo Fisher Scientific) was added to each well containing or not the SRC inhibitor eCF506 (100 nM, Medchem express) and in other cases the EphB inhibitors EPHB2i (ALW-II-49-7, 200 nM) and the pan-EPH inhibitor pan-EPHi (ALW-II-41-27, 100 nM, Medchem express).

Techniques: Derivative Assay, Cell Culture, MANN-WHITNEY, Activity Assay, Phospho-proteomics, Western Blot, Isolation

Journal: bioRxiv

Article Title: Slap restricts oncogenic Src-family kinase signaling to maintain colonic epithelial homeostasis

doi: 10.64898/2026.01.05.697659

Figure Lengend Snippet: (A) Slap-dependent colonic organoid development. Representative images of colon-derived organoids from Slap f/f and Slap f/f Villin-Cre ERT2 mice cultured in Matrigel for 2 days. Quantification of organoid number and area is shown as mean ± SEM from 100-200 organoids per mouse (n = 8 mice per group). ****p<0.0001 Mann–Whitney test. (B) Intestinal Slap deletion increases SFK activity and global protein tyrosine phosphorylation in colonic crypts. Representative immunoblots (right) and quantification (left) of phospho-SFK (pSRC) and total phospho-tyrosine levels in lysates from isolated colonic crypts of the indicated genotypes. Data are presented as mean ± SEM from n = 4-5 mice per group. *p<0.05; **p<0.001 t test. (C) Slap-dependent colonic organoid expansion requires SFK activity. Representative images and quantification of colon-derived organoids from Slap f/f and Slap f/f Villin-Cre ERT2 mice cultured for 2 days in the presence or absence of the SRC-family kinase inhibitor eCF506 (100 nM). Data are shown as mean ± SEM from 100-200 organoids per mouse, n = 4 mice per group. ***p<0.001, ****p<0.0001 Mann–Whitney test.

Article Snippet: Inhibitors used in this study: SRCi eCF506 (MedChemExpress, #HY-112096), EPHB2 inhibitor ALW-II-49-7 (EPHB2i, MedChemExpress, #HY-18833), Pan Eph inhibitor ALW-II-41-27 (pan-EPHi, MedCehmExpress, # HY-18007).

Techniques: Derivative Assay, Cell Culture, MANN-WHITNEY, Activity Assay, Phospho-proteomics, Western Blot, Isolation