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Characterization of organoid cultures and age differences in IESC phenotype: Organoid cultures derived from adult female (a-c) and male rats (d-f). Representative bright field images (n = 6) of IESC, plated in matrigel and cultured for 4–6 days (a,d). Scale bar: 25 μm. Immunohistochemistry (n = 6) for the intestinal stem cell marker Lgr5+, the epithelial cell marker NaKATPAse-α, and the cellular proliferation marker Ki67 all at 6 div in females (b,c) and males (e,f). Scale bar: 22 μm. Representative images (n = 5) for the Wnt signaling marker, Wnt3a (g) and its inhibitor, <t>DKK1</t> (h), and the senescence marker <t>γH2AX</t> (i) in organoids derived from adult and middle-aged males and females. Scale bar: 32 μm. Histogram depicting mean (±SEM) proportion of cells positive for Wnt3a (n = 5) (j), DKK1 (n = 5) (k) and γH2AX (n = 5) (l). Two-way ANOVA followed by Tukey-multiple comparisons test. *: p < 0.05.
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Characterization of organoid cultures and age differences in IESC phenotype: Organoid cultures derived from adult female (a-c) and male rats (d-f). Representative bright field images (n = 6) of IESC, plated in matrigel and cultured for 4–6 days (a,d). Scale bar: 25 μm. Immunohistochemistry (n = 6) for the intestinal stem cell marker Lgr5+, the epithelial cell marker NaKATPAse-α, and the cellular proliferation marker Ki67 all at 6 div in females (b,c) and males (e,f). Scale bar: 22 μm. Representative images (n = 5) for the Wnt signaling marker, Wnt3a (g) and its inhibitor, <t>DKK1</t> (h), and the senescence marker <t>γH2AX</t> (i) in organoids derived from adult and middle-aged males and females. Scale bar: 32 μm. Histogram depicting mean (±SEM) proportion of cells positive for Wnt3a (n = 5) (j), DKK1 (n = 5) (k) and γH2AX (n = 5) (l). Two-way ANOVA followed by Tukey-multiple comparisons test. *: p < 0.05.
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Characterization of organoid cultures and age differences in IESC phenotype: Organoid cultures derived from adult female (a-c) and male rats (d-f). Representative bright field images (n = 6) of IESC, plated in matrigel and cultured for 4–6 days (a,d). Scale bar: 25 μm. Immunohistochemistry (n = 6) for the intestinal stem cell marker Lgr5+, the epithelial cell marker NaKATPAse-α, and the cellular proliferation marker Ki67 all at 6 div in females (b,c) and males (e,f). Scale bar: 22 μm. Representative images (n = 5) for the Wnt signaling marker, Wnt3a (g) and its inhibitor, <t>DKK1</t> (h), and the senescence marker <t>γH2AX</t> (i) in organoids derived from adult and middle-aged males and females. Scale bar: 32 μm. Histogram depicting mean (±SEM) proportion of cells positive for Wnt3a (n = 5) (j), DKK1 (n = 5) (k) and γH2AX (n = 5) (l). Two-way ANOVA followed by Tukey-multiple comparisons test. *: p < 0.05.
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Santa Cruz Biotechnology anti dkk 1 antibody
Characterization of organoid cultures and age differences in IESC phenotype: Organoid cultures derived from adult female (a-c) and male rats (d-f). Representative bright field images (n = 6) of IESC, plated in matrigel and cultured for 4–6 days (a,d). Scale bar: 25 μm. Immunohistochemistry (n = 6) for the intestinal stem cell marker Lgr5+, the epithelial cell marker NaKATPAse-α, and the cellular proliferation marker Ki67 all at 6 div in females (b,c) and males (e,f). Scale bar: 22 μm. Representative images (n = 5) for the Wnt signaling marker, Wnt3a (g) and its inhibitor, <t>DKK1</t> (h), and the senescence marker <t>γH2AX</t> (i) in organoids derived from adult and middle-aged males and females. Scale bar: 32 μm. Histogram depicting mean (±SEM) proportion of cells positive for Wnt3a (n = 5) (j), DKK1 (n = 5) (k) and γH2AX (n = 5) (l). Two-way ANOVA followed by Tukey-multiple comparisons test. *: p < 0.05.
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Characterization of organoid cultures and age differences in IESC phenotype: Organoid cultures derived from adult female (a-c) and male rats (d-f). Representative bright field images (n = 6) of IESC, plated in matrigel and cultured for 4–6 days (a,d). Scale bar: 25 μm. Immunohistochemistry (n = 6) for the intestinal stem cell marker Lgr5+, the epithelial cell marker NaKATPAse-α, and the cellular proliferation marker Ki67 all at 6 div in females (b,c) and males (e,f). Scale bar: 22 μm. Representative images (n = 5) for the Wnt signaling marker, Wnt3a (g) and its inhibitor, DKK1 (h), and the senescence marker γH2AX (i) in organoids derived from adult and middle-aged males and females. Scale bar: 32 μm. Histogram depicting mean (±SEM) proportion of cells positive for Wnt3a (n = 5) (j), DKK1 (n = 5) (k) and γH2AX (n = 5) (l). Two-way ANOVA followed by Tukey-multiple comparisons test. *: p < 0.05.

Journal: Brain, behavior, and immunity

Article Title: Intestinal epithelial stem cell transplants as a novel therapy for cerebrovascular stroke

doi: 10.1016/j.bbi.2022.10.015

Figure Lengend Snippet: Characterization of organoid cultures and age differences in IESC phenotype: Organoid cultures derived from adult female (a-c) and male rats (d-f). Representative bright field images (n = 6) of IESC, plated in matrigel and cultured for 4–6 days (a,d). Scale bar: 25 μm. Immunohistochemistry (n = 6) for the intestinal stem cell marker Lgr5+, the epithelial cell marker NaKATPAse-α, and the cellular proliferation marker Ki67 all at 6 div in females (b,c) and males (e,f). Scale bar: 22 μm. Representative images (n = 5) for the Wnt signaling marker, Wnt3a (g) and its inhibitor, DKK1 (h), and the senescence marker γH2AX (i) in organoids derived from adult and middle-aged males and females. Scale bar: 32 μm. Histogram depicting mean (±SEM) proportion of cells positive for Wnt3a (n = 5) (j), DKK1 (n = 5) (k) and γH2AX (n = 5) (l). Two-way ANOVA followed by Tukey-multiple comparisons test. *: p < 0.05.

Article Snippet: Sections were incubated overnight at room temperature with primary antibodies (Lgr5+ (anti-rabbit)- 1:500ul, Novus Biological sciences, NLS1236; NaK-ATPase-alpha (anti-mouse)- 1: 500ul, Millipore, 05–369–25UG; Villin (anti-mouse)-1:250ul, Santa Cruz, SC-58897; Ki-67 (anti-rabbit)- 1:250ul, Abcam,ab16667; ZO-1, (anti-rabbit)- 1: 250ul, ThermoFisher Scientific, 691–7300; Wnt3 (anti-rabbit)- 1:250ul, Abcam, ab219412; DKK1 (anti-mouse-1:250ul, Santa Cruz, SC-374574; γH2AX (anti-rabbit)-1:250ul, Abcam, ab81299) for overnight at 4°.

Techniques: Derivative Assay, Cell Culture, Immunohistochemistry, Marker