mouse: math1-gfp  (Novartis)

Journal: Molecular and Cellular Biology

Article Title: Transcriptional Regulation of Math1 by Aryl Hydrocarbon Receptor: Effect on Math1 + Progenitor Cells in Mouse Small Intestine

doi: 10.1080/10985549.2022.2160610

Figure Lengend Snippet: AhR activation by β-NF in the presence of γ-secretase inhibitor increases Math1 + progenitor cells in intestinal organoids. (A) Math1 -GFP positive cells (green circle) include all types of mature secretory cells and secretory progenitor cells. Ki67 positive cells (red circle) include stem cells and two types of progenitor cells (i.e., secretory progenitor cells, absorptive progenitor cells). Thus, the Math1 -GFP and Ki67 double-positive cells indicated by the arrow are considered to be proliferative Math1 -GFP-positive cells (secretory progenitor cells). (B) Immunofluorescent images for the intestinal organoids derived from Math1 -GFP mice ( Math1-GFP +/+ ). The organoids were treated with vehicle, β-NF (β-naphthoflavone, 50 nM), DAPT (10 µM) followed by vehicle, and DAPT (10 µM) followed by β-NF (50 nM). Green, GFP; Red, Ki67; Blue, DAPI. Yellow arrows indicate the double-stained cells by GFP and Ki67. Bar, 30 µm. (C and D) The frequency of the organoids classified by the number of total Math1 -GFP positive cells ( Math1 -GFP positive/Ki67 positive cells and Math1 -GFP-positive/Ki67-negative) and single Math1 -GFP-positive cells ( Math1 -GFP positive/Ki67-negative). The number of Math1 -GFP positive cells was counted in each organoid and normalized by total cell number (DAPI-positive cell number). The frequency of evaluation values (less than 0.1 or more than 0.1) was compared between the organoids treated with vehicle and β-NF (C) and DAPT followed by vehicle and DAPT followed by β-NF (D). (E and F) The frequency of the organoids classified by the number of total Ki67 positive cells (Ki67-positive/ Math1 -GFP-positive and Ki67-positive/ Math1 -GFP negative), and single Ki67-positive cells (Ki67-positive/ Math1 -GFP-negative). The number of Ki67-positive cells was counted in each organoid and normalized by total cell number. The frequency of evaluation values (less than 0.3 or less than 0.5, and more than 0.3 or more than 0.5) was compared between the organoids treated with vehicle and β-NF (E) and DAPT followed by vehicle and DAPT followed by β-NF (F). (G and H) The frequency of the organoids classified by the number of Math1 -GFP and Ki67 double-positive cells. The number of double-positive cells was counted in each organoid and normalized by the total cell number. The evaluation values were classified as 0–0.01, 0.01–0.03, 0.03–0.05, 0.05–0.07, 0.07–0.09, 0.09–0.11, and more than 0.11. The frequency of evaluation values (less than 0.01 and more than 0.01) was compared between the organoids treated with vehicle and β-NF (G, left panel) and DAPT followed by vehicle and DAPT followed by β-NF (H, left panel). The distributions of values are presented in the right panels. At least 25 organoids in each treatment group were examined in each experiment; four or five individual experiments were performed. Values are mean ± SD. Two-way ANOVA was performed to analyze the data, where one factor was the treatment condition (vehicle and β-NF, or DAPT plus vehicle and DAPT plus β-NF), and the other factor was the indicated cell number (GFP-positive cell number, Ki67-positive cell number, or GFP and Ki67 double-positive cell number). The main effect of each factor was tested as well as the interaction within both factors.

Article Snippet: The Vil-Cre (No. 004586), AhR-Flox (No. 006203), and Math1-GFP (No. 013593) mouse lines were obtained from the Jackson Laboratory.

Techniques: Activation Assay, Derivative Assay, Staining

Journal: Molecular and Cellular Biology

Article Title: Transcriptional Regulation of Math1 by Aryl Hydrocarbon Receptor: Effect on Math1 + Progenitor Cells in Mouse Small Intestine

doi: 10.1080/10985549.2022.2160610

Figure Lengend Snippet: Sequential treatment with β-NF and γ-secretase inhibitor increases Math1 + progenitor cells in mouse small intestines. (A) Schema for treatment of mice. Math1-GFP mice ( Math1-GFP +/+ ) were treated with one dose of DBZ (30 µmol/kg BW, i.p.) followed by three doses of β-NF (β-naphthoflavone, 40 mg/kg BW, i.p.) or vehicle treatment. The samples were collected 24 h after the last treatment. (B) Expression level of Hes1 mRNA in the isolated crypts derived from the mice administrated one dose of DBZ (30 µmol/kg BW, i.p.) ( n = 3) or vehicle ( n = 3). The crypts were collected 24 h after treatment. Gene expression was normalized by Actb , and the expression level in the vehicle control was set as 1. (C) Body weights of mice before (pre) and after (post) the sequential treatment. (D) The immunofluorescent images for the small intestines of Math1-GFP mice ( Math1-GFP +/+ ). Green, GFP; Red; Ki67; Blue, DAPI. White and yellow arrows indicate the single- and the double-stained cells, respectively, by GFP and Ki67. Bar 50 µm. (E) The number of Math1 -GFP positive cells (total: Math1 -GFP-positive/Ki67-positive and Math1 -GFP-positive/Ki67-negative cells, single: Math1 -GFP-positive/Ki67-negative cells), Ki67-positive cells (total: Ki67-positive/ Math1 -GFP-positive and Ki67-positive/ Math1 -GFP-negative cells, single: Ki67-positive/ Math1 -GFP-negative cells), and Math1 -GFP/Ki67 double-positive cells in the intestinal crypts of the mice treated by DBZ plus vehicle ( n = 9) or DBZ plus β-NF ( n = 9). The cell number was normalized by total DAPI-positive cells in the crypts. At least 50 of well-oriented crypts were analyzed per section. Values are mean ± SD. * P < 0.05 using Student’s t test.

Article Snippet: The Vil-Cre (No. 004586), AhR-Flox (No. 006203), and Math1-GFP (No. 013593) mouse lines were obtained from the Jackson Laboratory.

Techniques: Expressing, Isolation, Derivative Assay, Gene Expression, Control, Staining

Journal: STAR Protocols

Article Title: Protocol to photoactivate adipose-derived stem cell differentiation using a tightly-focused femtosecond laser

doi: 10.1016/j.xpro.2022.101574

Figure Lengend Snippet:

Article Snippet: Mouse: Math1-GFP (male mice over 8 weeks old, female mice aged 4–6 weeks) , Novartis , N/A.

Techniques: Recombinant, In Vivo, Isolation, Plasmid Preparation, Software, Cell Culture, Microscopy