Journal: Advanced Science

Article Title: Stanniocalcin‐1 Promotes PARP1‐Dependent Cell Death via JNK Activation in Colitis

doi: 10.1002/advs.202304123

Figure Lengend Snippet: Stc1 deficiency alleviates DSS‐induced acute murine colitis. A) Body weight loss of WT, Stc1 INT‐KO , WT+DSS, and Stc1 INT‐KO +DSS mice. WT, n = 6; Stc1 INT‐KO , n = 7; WT+DSS, n = 8; Stc1 INT‐KO +DSS, n = 8. (B, C) Mice colon length and representative images of colons. WT, n = 6; Stc1 INT‐KO , n = 7; WT+DSS, n = 8; Stc1 INT‐KO +DSS, n = 8. D) DAI scores were used in the evaluation of colitis severity. WT, n = 6; Stc1 INT‐KO , n = 7; WT+DSS, n = 8; Stc1 INT‐KO +DSS, n = 8. (E, F) Histological scores and representative colonic histopathological images. WT, n = 6; Stc1 INT‐KO , n = 7; WT+DSS, n = 8; Stc1 INT‐KO +DSS, n = 8. Scale bars: 200 µm. (G, H) Representative colonoscopy images and endoscopic colitis scores were used to evaluate colitis severity observed under murine colonoscopy. WT, n = 3; Stc1 INT‐KO , n = 3; WT+DSS, n = 6; Stc1 INT‐KO +DSS, n = 6. I) TEM detected the deformation of mitochondria (marked by red arrows) in mice colonic epithelial cells. Scale bars: 1 µm. (J, K) Representative IF images of γ‐H2AX‐ J) and PAR‐stained K) mice colon sections. Scale bars: 100 µm. (L) The expression level of pro‐inflammatory cytokine mRNA in mice colonic tissues was detected via qPCR. WT, n = 5; Stc1 INT‐KO , n = 5; WT+DSS, n = 5; Stc1 INT‐KO +DSS, n = 5. M,N) The expression level of IL‐6 and TNF‐α protein in mice serum was detected via multiELISA. WT, n = 4; Stc1 INT‐KO , n = 4; WT+DSS, n = 4; Stc1 INT‐KO +DSS, n = 4. Data were expressed as mean ± SEM. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Fresh mice colonic tissues were harvested and made suitable for observation using TEM (HITACHI HT7800, Japan).

Techniques: Staining, Expressing

Journal: Advanced Science

Article Title: Stanniocalcin‐1 Promotes PARP1‐Dependent Cell Death via JNK Activation in Colitis

doi: 10.1002/advs.202304123

Figure Lengend Snippet: PARP1 activates JNK pathway via PARP1‐JNK interaction in an oxidate stress‐induced inflammatory environment. A) The subcellular distribution of PARP1 and JNK protein in NCM460 and Caco2 cell lines was detected by confocal microscopy. Scale bars: 10 µm. B,C) Anti‐PARP1 B) and anti‐JNK C) antibodies were respectively applied in the Co‐IP of STC1 OE NCM460 cells. D) His pull‐down assays of purified recombinant human His‐PARP1 and JNK protein. E) After the application of PARP inhibitor PJ34 (5µM, 24 h), the expression of JNK pathway protein in NCM460 and Caco2 cells was detected via western blot. F) The expression level of JNK pathway protein in the colonic tissues of WT, Stc1 INT‐KO , WT+DSS, and Stc1 INT‐KO +DSS mice was detected via western blot. G) The expression level of JNK pathway protein in H 2 O 2 ‐treated STC1 OE and STC1 KO cells were detected via western blot.

Article Snippet: Fresh mice colonic tissues were harvested and made suitable for observation using TEM (HITACHI HT7800, Japan).

Techniques: Confocal Microscopy, Co-Immunoprecipitation Assay, Purification, Recombinant, Expressing, Western Blot

Journal: Advanced Science

Article Title: Stanniocalcin‐1 Promotes PARP1‐Dependent Cell Death via JNK Activation in Colitis

doi: 10.1002/advs.202304123

Figure Lengend Snippet: Restoring Stc1 and Parp1 in vivo aggravates DSS‐induced mice colitis. A) Body weight loss of DSS‐treated Stc1 INT‐KO mice after peritoneal injection of Stc1 ‐overexpressing, Parp1 ‐overexpressing, or control AAV. Stc1 INT‐KO +DSS+Control AAV, n = 6; Stc1 INT‐KO +DSS+ Stc1 AAV, n = 6; Stc1 INT‐KO +DSS+ Parp1 AAV, n = 6. (B, C) Mice colon length and representative images of colons. Stc1 INT‐KO +DSS+Control AAV, n = 6; Stc1 INT‐KO +DSS+ Stc1 AAV, n = 6; Stc1 INT‐KO +DSS+ Parp1 AAV, n = 6. D) DAI score was used in the evaluation of colitis severity after AAV injection. Stc1 INT‐KO +DSS+Control AAV, n = 6; Stc1 INT‐KO +DSS+ Stc1 AAV, n = 6; Stc1 INT‐KO +DSS+ Parp1 AAV, n = 6. (E, F) Representative colonic histopathological images and histological scores. Stc1 INT‐KO +DSS+Control AAV, n = 6; Stc1 INT‐KO +DSS+ Stc1 AAV, n = 6; Stc1 INT‐KO +DSS+ Parp1 AAV, n = 6. Scale bars: 200 µm. G) Representative colonoscopy images were captured with murine colonoscopy. (H, I) Representative IF images of γ‐H2AX‐ H) and PAR‐stained I) mice colon sections. Scale bars: 100 µm. J) The expression level of pro‐inflammatory cytokine mRNA in mice colonic tissues was detected via qPCR. Stc1 INT‐KO +DSS+Control AAV, n = 4; Stc1 INT‐KO +DSS+ Stc1 AAV, n = 4; Stc1 INT‐KO +DSS+ Parp1 AAV, n = 4. K) The expression level of JNK pathway protein in mice colonic tissues was detected via western blot. Data were expressed as mean ± SEM. * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: Fresh mice colonic tissues were harvested and made suitable for observation using TEM (HITACHI HT7800, Japan).

Techniques: In Vivo, Injection, Control, Staining, Expressing, Western Blot