Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: LTH-sEV promotes LSEC capillarization in HFD mice and aggravates the progression of MASLD. ( A ) NTA analysis of LTH-sEV; ( B ) TEM images of LTH-sEV, scale bar = 100 nm; ( C ) Western blot analysis of CD9, CD63, Alix, TSG101, and calnexin protein expression in hepatocyte (HepG2) and LTH-sEV; ( D ) Schematic diagram of LTH-sEV injection into mice on HFD and NCD diet; ( E ) In vivo imaging detection of DiR-labeled LTH-sEV in mice; ( F ) SEM images and porosity of hepatic sinusoidal in NCD-fed mice treated with PBS, LTH-sEV, HFD-fed mice treated with PBS, LTH-sEV, n = 6, scale bar = 500 nm; ( G ) Immunohistochemistry images of CD31, Ang-2 in each group, scale bar = 50 μm, n = 6; ( H ) ELISA analysis of serum Ang-2 in each group, n = 6; ( I ) Western blot analysis of Ang-2 expression in each group, n = 3; ( J ) H&E staining, immunohistochemistry images of α-SMA, Sirius red staining in each group, scale bar = 50 μm, n = 6; ( K ) Serum ALT, AST expression in each group, n = 6; ( L ) qRT-PCR analysis of IL-1β, IL-6, and TNFα expression in each group, n = 6; ( M ) ELISA analysis of IL-1β, IL-6, and TNFα level in mice liver or serum, n = 6. Compared with HFD or NCD group, ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ns, no significance.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Western Blot, Expressing, Injection, In Vivo Imaging, Labeling, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Staining, Quantitative RT-PCR

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: OGT-enriched Hepatocyte-derived Extracellular Vesicles Promote Capillarization of Liver Sinusoidal Endothelial Cells in Metabolic Dysfunction-associated Steatotic Liver Disease

doi: 10.1016/j.jcmgh.2025.101721

Figure Lengend Snippet: OGT level in serum and serum sEV is positively associated with MASLD. ( A ) ELISA analysis of serum OGT expression in healthy individuals and patients with MASLD, n = 50; ( B ) ROC curve of serum OGT, AUC = .6832, P = .0016, Compared with Healthy group, ∗∗ P < .01; ( C ) Correlation analysis of serum OGT and ALT, r = .2154, P = .0438; ( D ) Correlation analysis of serum OGT and AST, r = .2493, P = .0171; ( E ) Correlation analysis of serum OGT and TG, r = .2541, P = .0197; ( F ) Immunohistochemical images of OGT expression in liver of healthy individuals and patients with MASLD; ( G ) Schematic diagram of the collection of Healthy-sEV and MASLD-sEV; ( H–I ) NTA and TEM detection of Healthy-sEV and MASLD-sEV, scale bar = 100 nm; ( J ) Western blot analysis of CD9, CD63, Alix and TSG101 protein expression in Healthy-sEV and MASLD-sEV; ( K ) Western blot analysis of OGT protein expression in Healthy-sEV and MASLD-sEV, n = 3; ( L ) ELISA analysis of of OGT in Healthy-sEV and MASLD-sEV, n = 50, Compared with Healthy-sEV group, ∗∗ P < .01; ( M ) ROC curve of OGT in serum sEV, AUC = .7228, P = .0001; ( N ) Correlation analysis of sEV-OGT and serum ALT, r = .2395, P = .0246; ( O ) Correlation analysis of sEV-OGT and serum AST, r = .2493, P = .0171; ( P ) Correlation analysis of sEV-OGT and Fib-4 score (Fib-4 = age × AST/PLT × √ALT), r = .3100, P = .0113.

Article Snippet: After blocking with 5% skim milk for 1 hour to eliminate nonspecific binding, the membrane was incubated overnight at 4°C with primary antibodies: β-actin (ABclonal, AC026, Rabbit), V5-Tag (ABclonal, AE101, Rabbit), TSG101 (Bioworld, BS91381, Rabbit), Calnexin (Bioworld, BS1438, Rabbit), CD9 (Proteintech, 60232-1-Ig, Mouse), CD63 (Abcam, ab271286, Rabbit), Ang-2 (Abcam, ab155106, Rabbit), CD31 (Proteintech, 11265-1-AP, Rabbit), OGT (Proteintech, 11576-2-AP, Rabbit), O-GlcNac (CST, #9875, Mouse), and HNF1α (Proteintech, 22426-1-AP, Rabbit).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemical staining, Western Blot

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of estrogen treatment on S100A8 expression in endometrium and epithelium morphology. ( a ) S100A8 immunohistochemical staining before and after estrogen treatment (1d/7d); black arrows indicate S100A8-positive immune cells (S100A8-PICs).The blue arrow indicates the release of S100A8 from S100A8-PICs. Scale bar is 100 or 400 μm. ( b ) Rat endometrial hematoxylin–eosin staining before and after estrogen treatment (1d/7d); red arrows show disrupted cell junctions. Scale bar is 100 μm. ( c ) S100A8-PICs number in the endometrium (1/mm 2 ) before and 1 d/7d after estrogen treatment (n = 18, from six rats, the number was counted from three immunohistochemical images of each sample , mean ± SEM). ( d ) S100A8 expression in the epithelium before and 1 d/7d after estrogen treatment (n = 18, from six rats, the expression level was measured from three immunohistochemical images of each sample, mean ± SEM). * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of uterine cavity administration of S100A8 on the distribution of S100A8-PICs and endometrial morphology. ( a ) Distribution of S100A8-positive cells in the endometrium of four different groups with estrogen treatment (control, injury, P407, and S100A8 + P407 groups). Black arrows show S100A8-PICs. S100A8-PICs were distributed around the glands and blood vessels in the control group. S100A8-PICs in the site of injury or adhesion in the injury group and in the P407 group. S100A8-PICs reverse migrated to the glands or blood vessels in the S100A8 + P407 group. Scale bar is 100 μm or 1 mm. ( b ) Endometrial thickness in the four groups (n = 15, from 5 rats; 3 position was measured randomly in an image of each rat, mean ± SEM). ( c ) The number of endometrial glands in each group (n = 10, from 10 rats, glands were counted from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). ( d ) The interior wall integrity of uterine cavity in each group (n = 10, from 10 rats, data was measured from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). ( e ) The number of endometrial crypts in each group (n = 10, from 10 rats, crypts were counted from a uterine horn cross-section image at 40 × of each rat, mean ± SEM). * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Control

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on the distribution of Ki-67-positive proliferating cells in the uteri and the proliferation of endometrial cells. ( a ) Ki-67-positive proliferating cells were distributed in the endometrium or uterine interior wall in all groups (control, injury, P407, and S100A8 + P407 groups). Scale bar is 100 or 400 μm. ( b ) Number of Ki-67-positive cells in the endometrium of each group (n = 20, from ten rats, the number was counted from two immunohistochemical images of each sample, mean ± SEM). ( c ) Number of Ki-67-positive cells in the uterine interior wall of each group (n = 20, from ten rats, the number was counted from two immunohistochemical images of each sample, mean ± SEM). ( d ) Effects of S100A8 and its receptor inhibitor (FPS-ZM1) on the proliferation of cultured endometrial cells in vitro (n = 5, mean ± SEM). Scale bar is 100 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Control, Immunohistochemical staining, Cell Culture, In Vitro

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on tight junctions of endometrial epithelium or endometrial cells. ( a ) Immunohistochemical staining showing Claudin-1 distribution in the endometrium of different groups (control, injury, P407, and P407 + S100A8 groups); black arrows show Claudin-1 accumulated below the cell membrane of the lateral contact zone between adjacent cells and red arrows show cells expressing Claudin-1 in the stroma. Scale bar is 100 μm. ( b ) Claudin-1 expression in the endometrium of different groups (n = 10, mean ± SEM). ( c ) Western blot showing ZO-1 expression in endometrial cells of three different groups (control, S100A8, and S100A8 + FPS-ZM1 groups, n = 3, mean ± SEM). Actin was used as the loading control. The full-length bands can be found in supplementary Fig. 1. The bands of ZO-1 and actin cropped from different parts of the same gel. ( d ) Immunofluorescence showing the co-expression of Claudin-1 (red) and ZO-1 (green) in the endometrial cells of the three groups (n = 5, mean ± SEM). The nucleus is stained blue. The scale bar is 200 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Immunohistochemical staining, Staining, Control, Membrane, Expressing, Western Blot, Immunofluorescence

Journal: Scientific Reports

Article Title: S100A8 regulated by estrogen improves injured endometrial epithelium reconstruction by promoting tight junction formation and stromal cell transformation

doi: 10.1038/s41598-025-08530-0

Figure Lengend Snippet: Effects of S100A8 on endometrial “double-featured” cells. ( a ) Vimentin (red) and ZO-1 (green) immunofluorescent double staining of cultured endometrial cells. Scale bar is 200 μm. ( b ) Western blot showing CK-18 and vimentin expression in endometrial cells from each group (control, S100A8, and FPS-ZM1 groups; n = 3, mean ± SEM). The full-length bands can be found in supplementary Fig. 1. Actin was used as the loading control. As the target proteins have a similar molecular weight to that of the loading control, the bands of CK18/vimentin and actin were cropped from different gels. ( c ) Immunofluorescence showing vimentin expression in endometrial cells of each group (n = 5, mean ± SEM). Scale bar is 200 μm. * Results with P values < 0.05 were considered significant. ** Results with P values < 0.01 were considered very significant.

Article Snippet: The cultured cells were randomly divided into three groups according to the experimental design: the control, S100A8 (0.5 μg/mL, HY- P71275 ; MCE, Shanghai, China), and S100A8 + FPS-ZM 1 (5 μM, HY-19370; MCE, Shanghai, China) groups.

Techniques: Double Staining, Cell Culture, Western Blot, Expressing, Control, Molecular Weight, Immunofluorescence