(A and B) Image of Ki67‐positive cells (Red: Ki67, Blue: DAPI) and their quantification at the crypt base of proximal jejunum (A) or colon (B) of NIC-treated and untreated mice (A:3–4 mice per …
The quantification in immunostaining.
(A) Images of the H&E-stained crypt and villus and quantification of their length after or without NIC treatment (3–4 mice per group). (B) The quantification of the number of enterocytes per villus …
The quantification in staining.
(A) Crypts from the proximal small intestine were cultured with 10 μM, 1 μM, and 100 nM NIC or cotinine to allow ISCs to form organoid colonies; the control set contained no NIC or cotinine. …
The quantification of colonies in organoid assay.
ISCs and Paneth cells were isolated from the small intestine of Lgr5‐EGFP‐IRES‐CreERT2 mice (NIC-treated and untreated); 2×103 cells were co-cultured using medium without 10 μM CHIR99021. …
The quantification of colonies in organoid assay.
(A) Isolated ISCs were cultured in a medium with or without 1 μM NIC and 10 μM Mecamylamine (3 wells/group). (B) Isolated ISCs were cultured in a medium with or without 1 μM NIC and 3 μM Adiphenine …
Colony quantification and qRT-PCR.
Immunoblotting data with labeling.
Immunoblotting raw data.
(A) Violin plots for nAchR subunits in ISCs and Panth cells by the analysis of human gut single-cell datasets (Elmentaite et al., 2021). (B) The number of positive cells for each nAchR in ISCs and …
(A and B) Isolated ISCs cultured using a medium supplemented with or without 1 μM NIC combined with either (A) 1 μM H89 dihydrochloride (PKA inhibitor) or (B) 10 nM Gö 6983 (PKC inhibitor; 3 …
Colony quantification and qRT-PCR.
Immunoblotting data with labeling.
Immunoblotting raw data.
(A) Isolated ISCs cultured using a medium supplemented with or without 1 μM NIC combined with 5 nM Sotrastaurin (PKC inhibitor) (3 wells/group). (B) Immunoblotting analyses of p38, p-p38, p-S6, or …
The quantification of colonies in organoid assay.
Immunoblotting data with labeling.
Immunoblotting raw data.
(A) Isolated ISCs were cultured using a medium with or without 1 μM Nicotine and 5 nM K-975 (3 wells/group). (B) Isolated ISCs were cultured using a medium with or without 1 μM Nicotine and 1 μM …
The quantification of colonies in organoid assay.
(A) Schematic representation of the treatment showing daily injection of DBZ (1 mg/kg body weight) for 2 weeks. (B) Immunoblotting analysis of crypt lysate isolated from DBZ- and vehicle-treated …
The quantification in immunostaining.
Immunoblotting data with labeling.
Immunoblotting raw data.
(A and B) Immuno-stained Ki67-positive cells (A) (Red, Ki67; Blue, DAPI) and LgR5-GFP-positive cells (B) (Red, Olfm4; blue, DAPI) and quantification of their abundance in the colon of DBZ or …
The quantification in immunostaining.
(A) Schematic representation of Apcflox/flox; Lgr5-EGFP-IRES-CreERT2 (Lgr5CreERT2 Apcfl/fl)tumor initiation. Mice were treated with control or NIC more than 8 weeks before a single Tamoxifen …
The quantification of polyps and β-catenin positive lesions.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Mus musculus) | Lgr5-EGFP-IRES-CreERT2 mice | Jackson Laboratory | #008875 | |
Strain, strain background (Mus musculus) | Apc CKO mice | National Cancer Institute | #01XAA | |
Strain, strain background (Mus musculus) | Rosa26-CAG-lsl-tdTomato mice | Jackson Laboratory | #007909 | |
Antibody | rabbit monoclonal anti-Ki67 | Cell Signaling Technology | #12202 | IHC (1:200) |
Antibody | rabbit monoclonal anti-Olfm4 | Cell Signaling Technology | #39141 | IHC (1:400) |
Antibody | mouse monoclonal anti-GFP | Santa Cruz Biotechnology | #sc-9996 | IHC (1:50) |
Antibody | rabbit polyclonal anti-Lysozyme | Thermo Fisher Schientific | #PA5-16668 | IHC (1:50) |
Antibody | mouse monoclonal anti-chromogranin A | Santa Cruz Biotechnology | #sc-393941 | IHC (1:50) |
Antibody | rabbit polyclonal anti-β-catenin | Cell Signaling Technology | #9562 | IHC (1:200) |
Antibody | goat polyclonal anti-mouse IgG H&L (Alexa Fluor 488) | Abcam | #ab150113 | IHC (1:200) |
Antibody | mouse monoclonal anti-β-actin | Santa Cruz | #sc-47778 | WB (1:200) |
Antibody | mouse monoclonal anti-YAP | Santa Cruz | #sc-101199 | WB (1:200) |
Antibody | mouse monoclonal anti-TAZ | Santa Cruz | #sc-293183 | WB (1:200) |
Antibody | mouse monoclonal anti-α7-AchR | Santa Cruz | #sc-58607 | WB (1:200) |
Antibody | mouse monoclonal anti-Notch1 | Santa Cruz | #sc-376403 | WB (1:100) |
Antibody | mouse monoclonal anti-Jagged1 | Santa Cruz | #sc-390177 | WB (1:100) |
Antibody | mouse monoclonal anti-Jagged2 | Santa Cruz | #sc-515725 | WB (1:100) |
Antibody | mouse monoclonal anti-Hes5 | Santa Cruz | #sc-293445 | WB (1:200) |
Antibody | mouse monoclonal anti-p38 | Santa Cruz | #sc-81621 | WB (1:200) |
Antibody | mouse monoclonal anti-phospho-p38 | Santa Cruz | #sc-166182 | WB (1:100) |
Antibody | rabbit monoclonal anti-S6 | Cell Signaling | #2217 | WB (1:1000) |
Antibody | rabbit monoclonal anti-phospho-S6 Ser235/236 | Cell Signaling | #4858 | WB (1:2000) |
Antibody | anti-Mouse IgG, HRP-Linked Whole Ab Sheep | Cytiva | NA931 | WB (1:5000) |
Antibody | anti-Rabbit IgG, HRP-Linked Whole Ab Donkey | Cytiva | NA934 | WB (1:5000) |
Antibody | rat monoclonal APC-conjugated anti-mouse CD24 Antibody | Biolegend | #101814 | FCY (1:500) |
Commercial assay or kit | TSA Plus Cyanine 3 System | Akoya Biosciences | #NEL744001KT | |
Chemical compound, drug | Mecamylamine | Cayman Chemical | #14602 | |
Chemical compound, drug | Adiphenine hydrochloride | MedChemExpress | #HY-B0379A | |
Chemical compound, drug | PNU282987 | MedChemExpress and Cayman Chemical | #17424 #HY-12560A | |
Chemical compound, drug | α-Bungarotoxin | R&D | #2133/1 | |
Chemical compound, drug | H-89 dihydrochloride | MedChemExpress | #HY-15979A | |
Chemical compound, drug | Gö 6983 | Cayman Chemical | #13311 | |
Chemical compound, drug | Sotrastaurin | MedChemExpress | #HY-10343 | |
Chemical compound, drug | K-975 | MedChemExpress | #HY-138565 | |
Chemical compound, drug | MK-0752 | MedChemExpress | #HY-10974 | |
Chemical compound, drug | Ingenol-3-angelate | Cayman Chemical | #16207 | |
Chemical compound, drug | AKT Inhibitor VIII | MedChemExpress | #HY-10355 | |
Chemical compound, drug | SB203580 | Tokyo Chemical Industry | #F0864 | |
Chemical compound, drug | Rapamycin | LKT Laboratories, Inc | #R0161 | |
Chemical compound, drug | Collagenase Type IV | Worthington Biochemical Corporation | #CLS4 | |
Chemical compound, drug | Valproic acid sodium salt | FUJIFILM Wako Pure Chemical Corporation | #2815/100 | |
Chemical compound, drug | [-]-Cotinine | Sigma-Aldrich | #C-016 | |
Chemical compound, drug | Nicotine hemisulfate salt | Sigma-Aldrich | #N1019 | |
Chemical compound, drug | DBZ (Dibenzazepine) | Cayman Chemical | #14627 | |
Chemical compound, drug | Tamoxifen | Cayman Chemical | #13258 | |
Chemical compound, drug | Mounting medium With DAPI Aqueous Fluoroshield | Abcam | #ab104139 | |
Chemical compound, drug | Can Get Signal Immunoreaction Enhancer Solution | TOYOBO | NKB-101 |