Figure 7.
TA cell death decreases R-spondin production in the crypt niche. (A) Scheme describing the generation of URI(+/+)Int; p53(Δ/Δ)Int and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice by crossing URI lox mouse, p53 lox mouse, and Villin-creERT2 mouse. (B) Kaplan–Meier curve of URI(+/+)Int; p53(+/+)Int, URI(+/+)Int; p53(Δ/Δ)Int, URI(Δ/Δ)Int; p53(+/+)Int and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice (n = 5, 8, 8, 8). (C) Representative pictures of H&E staining in intestines from URI(+/+)Int; p53(Δ/Δ)Int, URI(Δ/Δ)Int; p53(+/+)Int and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice at indicated time points of tamoxifen treatment. (D) Representative co-IF of R-spondin 1 and lysozyme in intestinal sections from URI(+/+)Int; p53(+/+)Int, URI(Δ/Δ)Int; p53(+/+)Int, URI(+/+)Int; p53(Δ/Δ)Int, and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice after 6 d of tamoxifen treatment. (E) qRT-PCR of Rspo1 and Rspo3 mRNA levels from intestinal tissue from URI(+/+)Int, URI(Δ/Δ)Int; p53(+/+)Int, and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice (n = 3). (F) Scheme depicting chemical inhibition of apoptosis and/or pyroptosis in URI lox mice. (G) Kaplan–Meier curve of URI(Δ/Δ)Int mice treated with vehicle (PBS; red solid line; n = 3), URI(Δ/Δ)Int mice treated with caspase 3 inhibitor (iCasp. 3; dark blue solid line; n = 3), URI(Δ/Δ)Int mice treated with caspase 1 inhibitor (iCasp. 1; grey solid line; n = 3) and URI(Δ/Δ)Int mice treated with both inhibitors (light blue solid line; n = 3). (H) Representative pictures of H&E staining from treated mice from F at the time of death (8–10 d). (I) Number of regenerative areas per optical field (76 mm2) observed in H. (J) Scheme depicting chemical inhibition of pyroptosis in URI(Δ/Δ)Int; p53(Δ/Δ)Int mice. (K) Kaplan–Meier curve of URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with vehicle (red solid line; n = 4) and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with caspase 1 inhibitor (green solid line; n = 4). (L) Representative pictures of H&E staining from URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated either with vehicle or caspase 1 inhibitor at the time of death (8–10 d). (M) Number of regenerative areas per optical field (76 mm2) observed in URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with vehicle and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with caspase 1 inhibitor at time of death (8–10 d; n = 4 mice per group). (N) qRT-PCR of Rspo1 and Rspo3 mRNA levels from URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated either with vehicle or caspase 1 and 3 inhibitors and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with caspase 1 inhibitor (n = 3). Data represent mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; Mantel–Cox, one-way ANOVA and Student’s t test. Scale bars represent 20 μm in D; and 100 µm in C, H, and L. H&E and IF are representative of at least three independent mice.

TA cell death decreases R-spondin production in the crypt niche. (A) Scheme describing the generation of URI(+/+)Int; p53(Δ/Δ)Int and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice by crossing URI lox mouse, p53 lox mouse, and Villin-creERT2 mouse. (B) Kaplan–Meier curve of URI(+/+)Int; p53(+/+)Int, URI(+/+)Int; p53(Δ/Δ)Int, URI(Δ/Δ)Int; p53(+/+)Int and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice (n = 5, 8, 8, 8). (C) Representative pictures of H&E staining in intestines from URI(+/+)Int; p53(Δ/Δ)Int, URI(Δ/Δ)Int; p53(+/+)Int and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice at indicated time points of tamoxifen treatment. (D) Representative co-IF of R-spondin 1 and lysozyme in intestinal sections from URI(+/+)Int; p53(+/+)Int, URI(Δ/Δ)Int; p53(+/+)Int, URI(+/+)Int; p53(Δ/Δ)Int, and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice after 6 d of tamoxifen treatment. (E) qRT-PCR of Rspo1 and Rspo3 mRNA levels from intestinal tissue from URI(+/+)Int, URI(Δ/Δ)Int; p53(+/+)Int, and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice (n = 3). (F) Scheme depicting chemical inhibition of apoptosis and/or pyroptosis in URI lox mice. (G) Kaplan–Meier curve of URI(Δ/Δ)Int mice treated with vehicle (PBS; red solid line; n = 3), URI(Δ/Δ)Int mice treated with caspase 3 inhibitor (iCasp. 3; dark blue solid line; n = 3), URI(Δ/Δ)Int mice treated with caspase 1 inhibitor (iCasp. 1; grey solid line; n = 3) and URI(Δ/Δ)Int mice treated with both inhibitors (light blue solid line; n = 3). (H) Representative pictures of H&E staining from treated mice from F at the time of death (8–10 d). (I) Number of regenerative areas per optical field (76 mm2) observed in H. (J) Scheme depicting chemical inhibition of pyroptosis in URI(Δ/Δ)Int; p53(Δ/Δ)Int mice. (K) Kaplan–Meier curve of URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with vehicle (red solid line; n = 4) and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with caspase 1 inhibitor (green solid line; n = 4). (L) Representative pictures of H&E staining from URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated either with vehicle or caspase 1 inhibitor at the time of death (8–10 d). (M) Number of regenerative areas per optical field (76 mm2) observed in URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with vehicle and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with caspase 1 inhibitor at time of death (8–10 d; n = 4 mice per group). (N) qRT-PCR of Rspo1 and Rspo3 mRNA levels from URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated either with vehicle or caspase 1 and 3 inhibitors and URI(Δ/Δ)Int; p53(Δ/Δ)Int mice treated with caspase 1 inhibitor (n = 3). Data represent mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; Mantel–Cox, one-way ANOVA and Student’s t test. Scale bars represent 20 μm in D; and 100 µm in C, H, and L. H&E and IF are representative of at least three independent mice.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal