Claudin 1 Antibody - #AF0127

Fig. 5: Flea bites affect neural function through the skin-brain axis. a GSEA result on upregulated genes from the mice skin transcriptome. Functions related to immunoglobulin and hemoglobin genes are significantly enriched. b Cytokine levels in mice skin presented in boxplot where red is Flea− (n = 6/12) and blue is Flea+ (n = 6/12). Two-sided t test was used. Asterisks indicate the significance. All p-values were below 0.001. c Cytokine levels in mice serum presented in boxplot where red is Flea− (n = 12) and blue is Flea+ (n = 12). Two-sided t test was used. Thick bars indicate the interquartile range (IQR) around the median, and whiskers represent 1.5 times the interquartile range (maxima: Q3 + 1.5 × IQR, minima: Q1 − 1.5 × IQR). Asterisks indicate the significance. All p-values were below 0.001. d Cytokine levels in mice PFC presented in boxplot where red is Flea− (n = 10) and blue is Flea+ (n = 10). Two-sided t test was used. Asterisks indicate the significance. All p-values were below 0.001. e Representative HE plots of the PFC. In the Flea+ group, cortical tissue shows evident cellular damage, with minor mononuclear cell infiltration (yellow arrow) at the injury site. Additionally, widespread nuclear pyknosis and condensation (red arrow) are observed in the parenchyma. The infected group exhibits a larger area of damage; Scale 50 µm. f Western blot analysis of Claudin1 markers of tight junction protein from the PFC brain region (n = 5 samples each group). g Comparison of Claudin1 in WB between the Flea− (red, n = 5) and Flea+ (blue, n = 5) groups of mice. Two-sided t test was used. Each black dot represents an individual. Asterisks indicates brain regions with significant differences. Data are presented as mean ± SEM. Source data are provided as a Source Data file. h Validation by qPCR of select BBB genes altered between Flea− (n = 3) and Flea+ (n = 3) group. Two-sided t test was used. Thick bars indicate the interquartile range (IQR) around the median, and whiskers represent 1.5 times the interquartile range (maxima: Q3 + 1.5 × IQR, minima: Q1 − 1.5 × IQR). Each black dot represents pooled sample of 3 decapitated individuals. Asterisks indicate the significance.

Figure 5. |FnEVs increase gut barrier leakage in experimental colitis models. (a) Representative images and ex vivo imaging with the intestine, liver, heart, spleen and kidney of mice. (b) Relative fluorescence intensity of translocated EGFP-labeled E.coli in every tissues. (c) Representative images of immunohistochemical stainings of ZO-1, claudin-1 and occludin in the colon on day 3 after colitis induction. Scale bar = 50 um.

Fig. 3. The mRNA level, protein expression level and Western blotting analysis of tight junction proteins in intestinal and lung tissues of AAI mice. A, Relative mRNA expression levels of Occludin, Claudin-1 and ZO1 in lung and gut tissues. B, Representative Western-blot images of Occludin, Claudin1, and ZO1 proteins in gut and lung tissues with β-Actin as the internal reference. Statistical significance was determined by Student's t-test.

Fig. 4. OBB protected intestinal epithelial barrier by modulating TJs proteins. Effect of OBB on the mRNA levels of mucin-1(A) and mucin-2 (B). (C) Representative Western blotting images of TJs protein, and the relative protein expressions were normalized to β-actin. (D-H) Changes in the relative protein expression levels of ZO-1, ZO-2, occludin, JAM-A, and claudin-1 were measured respectively. Data are shown as mean ± SEM (n = 3). # P < 0.05, ## P < 0.01 vs. Control group, * P < 0.05, ** P < 0.01 vs. DSS group, & P < 0.05, && P < 0.01 vs. BBR group.

Figure 2 The immunofluorescence of organoids with or without LPS stimulation. The images were captured by Opera Phenix™ high-content imaging system and compressed by Harmony. (A) The expression of Ki-67, LPS 100 μg/mL, 24 h. (B) The expression of ZO-1, LPS 100 μg/mL, 24 h. (C) The expression of occludins, LPS 100 μg/mL, 24 h. (D) The expression of claudin-1, LPS 100 μg/mL, 24 h. (E) Quantitative analysis of Ki-67 with different concentrations of LPS stimulated for 8 h or 24 h. (F) Quantitative analysis of ZO-1 with different concentrations of LPS stimulated for 8 h or 24 h. (G) Quantitative analysis of occludins with different concentrations of LPS stimulated for 8 h or 24 h. (H) Quantitative analysis of claudin-1 with different concentrations of LPS stimulated for 8 h or 24 h. Data are presented as means ± SD, compared with LPS 0 μg/mL 8 h or 24 h group accordingly using Student's t-tests. ∗ is used for 24 h. SD: Standard deviation; LPS: Lipopolysaccharide; ZO-1: Zonula occludens-1.

Figure 3. SIRT1 deacetylates and stabilizes β-TrCP1 protein to downregulate Snail1. (A) Caco2 cells stably expressing shRNAs against SIRT1 were subjected to Western blotting. (B) Caco2 cell lysates were subjected to immunoprecipitation (IP) with anti-β-TrCP1, using normal mouse or rabbit IgG as a control, followed by Western blot analyses. (C) Caco2 cells stably coexpressing β-TrCP1 and shRNAs against SIRT1 were subjected to Western blotting. (D) Caco2 cells treated with or without DSS were subjected to Western blotting. (E) HCT116 cells were treated with 4% DSS for 8 hours and the whole cell lysates were subjected to Western blotting. (F) Caco2 cells stably overexpressing SIRT1 were treated with 4% DSS for 8 hours. Whole-cell lysates were subjected to Western blotting. (G) Caco2 cells stably overexpressing β-TrCP1 were treated with 4% DSS for 8 hours. Whole-cell lysates were subjected to Western blotting. (H) Caco2 cells pretreated with 4% DSS for 8 hours or stably expressing shRNAs against SIRT1 were subjected to qPCR analyses for β-TrCP1. (I) To examine the effects of DSS on ubiquitination of β-TrCP1, 293FT cells were cotransfected with the relevant expression plasmids for 48 hours, followed by treatment with 4% DSS for 12 hours. Before harvesting, cells underwent treatment with 20 μM MG132 for 6 hours. Ubiquitination of β-TrCP1 was examined by denature-IP-western analyses. (J) The β-TrCP1 protein half-lives were measured by CHX assays in Caco2 cells pretreated with 4% DSS for 8 hours. (K and L) To examine the effects of SIRT1 on ubiquitination of β-TrCP1 and Snail1, 293FT cells were cotransfected with indicated expressing plasmids. Cells were treated with 20 μM MG132 for 6 hours before collection. Ubiquitination of Snail1 was examined by denature-IP-western analyses. (M) The β-TrCP1 and Snail1 protein half-lives were measured by CHX assays in Caco2 cells stably overexpressing SIRT1. (N and O) Caco2 cell lysates were subjected to IP with anti-SIRT1 (H) or anti-β-TrCP1 (I), using normal mouse or rabbit IgG as a control, followed by Western blot analyses. (P) HEK-293T cells cotransfected with His/Myc-β-TrCP1 and Flag-SIRT1 were subjected to IP and Western blotting analyses.

Fig. 6 Palmitoleic acid promotes intestinal tight junction integrity and reduces HFD-induced colon inflammation. 6 weeks old mice were fed with HFD for 12 weeks. Mice were treated with palmitoleic acid, oleic acid, or BSA as control via oral gavages for 6 weeks. The colon was collected for further analysis. (A) The level of occludin and claudin-1 by Western blotting. (B) Quantification of occludin level. (C) Quantification of claudin-1 level. (D) Macrophage infiltration by IHC for F4/80. (E) mRNA levels of MCP-1, TNF-α, IL-1β, and IL-6 by RT-PCR. n = 4 mice per group. The data are mean ± SEM (error bars). † P < 0.05, ††P < 0.01, †††P < 0.001, BSA vs palmitoleic acid; ‡ P < 0.05, ‡‡P < 0.01, ‡‡‡P < 0.001, oleic acid vs palmitoleic acid; §§§P < 0.001, BSA vs oleic acid.

Figure 7 Effect of djulis hull crude extract on (A) LPS in serum and (B) the protein expression of tight junctions in the colon of high-fat diet-induced hyperglycaemia. ND: normal diet; HFD: high-fat diet; HCE: high dosage of crude extract; LPS: lipopolysaccharide. Values represent the mean ± SEM (n = 6). The statistical methods used one-way ANOVA, and the values with different letters are significantly different at p < 0.05.

Fig. 4. Effects of AITC on tight junction-related protein in lung. AITC treatments on the modulation of tight junction-related protein expression. (A) E-cadherin, (B), ZO-1, (C) Occludin, (D) Claudin-1. Data are expressed in mean ± SEM (n = 5–7). Significant difference presented as different symbol. *p