Journal: Nutrients

Article Title: Synbiotic Supplementation Containing Whole Plant Sugar Cane Fibre and Probiotic Spores Potentiates Protective Synergistic Effects in Mouse Model of IBD

doi: 10.3390/nu11040818

Figure Lengend Snippet: Effect of B. coagulans spores, PSCF and synbiotic on immune markers in colon tissues and blood serum. Protein levels of cytokines including ( A ) IL-1α, ( B ) IL-1β, ( C ) IL-6, ( D ) IL-12, ( E ) TNF-α, ( F ) IFN-γ in proximal and distal colon explants as well as cytokine levels of ( G ) IL-1β, ( H ) IL-10, and ( I ) IL-12 in blood serum were analysed by Bio-plex. iNOS activity in colon tissues ( J ) measured by NOS activity assay and CRP levels in serum ( K ) by ELISA. Statistical significance among groups evaluated by one-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. DSS-colitic group and data expressed as mean ± SEM ( n = 3 per group).

Article Snippet: The levels of C-reactive protein (CRP) in serum from respective groups ( n = 3 samples/group) were analysed using Mouse C-Reactive Protein/CRP Quantikine Elisa kit (MCRP00, R and D Systems, Australia) following the manufacturer’s instructions.

Techniques: Activity Assay, Nos Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal for immunotherapy of cancer

Article Title: Endosialin-positive CAFs promote hepatocellular carcinoma progression by suppressing CD8 + T cell infiltration.

doi: 10.1136/jitc-2024-009111

Figure Lengend Snippet: Figure 4 Knockdown of endosialin promotes the expression and secretion of CXCL9/10 in cancer-associated fibroblasts (CAFs) by activating INF-γ-STAT1 pathway. (A) Chemokine array to show the levels of different chemokines in IFN-γ-stimulated endosialin knockdown or control HFL1 cells. (B) RT-qPCR to show the mRNA level of CXCL9/10 in endosialin knockdown or control HFL1 cells. (C) ELISA to show the protein level of CXCL9/10 in the culture medium of endosialin knockdown or control HFL1 cells. (D) RT-qPCR to show the mRNA level of CXCL9/10 in endosialin overexpressing or control LX-2 cells. (E) ELISA to show the protein level of CXCL9/10 in the culture medium of endosialin overexpressing or control LX-2 cells. (F) ELISA to show the level of CXCL9/10 in subcutaneous Hepa1-6 tumor tissues of WT and ENKO mice (n=3). (G) Fluorescent imaging to show the migrated 5(6)-Carboxyfluorescein diacetate N-succinimidyl ester (CFSE)-labeled T cells after co-culture with endosialin knockdown or control HFL1 cells in the presence of CXCL10 neutralizing antibody or control antibody (left) and quantification of the migrated T cells (right). Scale bar, 100 µm. (H) Western blot to detect the level of pSTAT1 in endosialin knockdown or control HFL1 cells. (I) IF staining to show the localization of STAT1 in endosialin knockdown or control HFL1 cells. Scale bar, 20 µm. (J) Western blot to detect the level of pSTAT1 in endosialin overexpressing or control LX-2 cells. (K) IF staining to show the localization of STAT1 in endosialin overexpressing or control LX-2 cells. Scale bar, 20 µm. Representative images are shown. Data are presented as the mean±SEM. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: To examine CXCL9/10 level in the tumor tissues of ENKO or WT mice, tumor tissues were isolated and prepared into a tissue homogenate by tissue fragmentation, and then examined using mouse CXCL9 (CUSABIO, #CSB- EL006252MO) and CXCL10 (CUSABIO, #CSBE08183m) ELISA kits according to the manufacturer’s instructions.

Techniques: Knockdown, Expressing, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Imaging, Labeling, Co-Culture Assay, Western Blot, Staining

Journal: Journal for immunotherapy of cancer

Article Title: Endosialin-positive CAFs promote hepatocellular carcinoma progression by suppressing CD8 + T cell infiltration.

doi: 10.1136/jitc-2024-009111

Figure Lengend Snippet: Figure 5 High endosialin expression is correlated with low CD8+ T infiltration in clinical hepatocellular carcinoma (HCC) tissues. (A) Immunohistochemistry (IHC) staining of endosialin in HCC tissues and adjacent normal tissues (n=5). Scale bar, 100 µm (top panel), 20 µm (bottom panel). (B) Dual immunofluorescent (IF) staining of HCC tissues to show the correlation between endosialin expression and CD8+ T cell infiltration (n=4). Scale bar, 50 µm. (C) RT-qPCR to show the mRNA level of CXCL9/10 in endosialin knockdown or control primary cancer-associated fibroblasts (CAFs. (D) ELISA to show the protein level of CXCL9/10 in the culture medium of endosialin knockdown or control primary CAFs. (E) Fluorescent imaging to show the migrated 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE)-labeled T cells after co-culture with endosialin knockdown or control primary CAFs at the presence of CXCL10 neutralizing antibody or control antibody (left) and quantification of the migrated T cells (right). Scale bar, 100 µm. (F) Western blot to detect the level of pSTAT1 in endosialin knockdown or control primary CAFs. (G) IF staining to show the localization of STAT1 in endosialin knockdown or control primary CAFs. Scale bar, 20 µm. Representative images are shown. Data are presented as the mean±SEM. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: To examine CXCL9/10 level in the tumor tissues of ENKO or WT mice, tumor tissues were isolated and prepared into a tissue homogenate by tissue fragmentation, and then examined using mouse CXCL9 (CUSABIO, #CSB- EL006252MO) and CXCL10 (CUSABIO, #CSBE08183m) ELISA kits according to the manufacturer’s instructions.

Techniques: Expressing, Immunohistochemistry, Staining, Quantitative RT-PCR, Knockdown, Control, Enzyme-linked Immunosorbent Assay, Imaging, Labeling, Co-Culture Assay, Western Blot

Journal: Journal for immunotherapy of cancer

Article Title: Endosialin-positive CAFs promote hepatocellular carcinoma progression by suppressing CD8 + T cell infiltration.

doi: 10.1136/jitc-2024-009111

Figure Lengend Snippet: Figure 6 Combination therapy with endosialin antibody and PD-1 antibody shows synergistic antitumor effect. (A) Schematic image to show the process of combination therapy. (B) Picture of isolated tumors (left) and tumor weight (right) after mice were sacrificed after treatment (n=8). (C) Growth curve of the tumors in different groups after treatment. (D) Flow cytometry to show the percentage of CD4+ T cells and CD8+ T cells in the tumor tissues of different groups after treatment (left panel) and quantification of the flow cytometry data (right panel) (n=6). (E) Graphic diagram to describe the function of endosialin, which can inhibit the secretion of CXCL9/10 by inhibiting IFN-γ/JAK/STAT1 pathway in CAFs, thus inhibit the infiltration of CD8+ T cells into tumor tissues; and endosialin antibody can enhance the anti-tumor effect of PD1 antibody by increasing the infiltration of CD8+ T cells. Representative images are shown. Data are presented as the mean±SEM. *p<0.05; **p<0.01; ***p<0.001; ns, not significant. CAFs, cancer-associated fibroblasts.

Article Snippet: To examine CXCL9/10 level in the tumor tissues of ENKO or WT mice, tumor tissues were isolated and prepared into a tissue homogenate by tissue fragmentation, and then examined using mouse CXCL9 (CUSABIO, #CSB- EL006252MO) and CXCL10 (CUSABIO, #CSBE08183m) ELISA kits according to the manufacturer’s instructions.

Techniques: Isolation, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Pathogenic Roles of CXCL10 in Experimental Autoimmune Prostatitis by Modulating Macrophage Chemotaxis and Cytokine Secretion

doi: 10.3389/fimmu.2021.706027

Figure Lengend Snippet: CXCL10 expression is increased in chronic prostatitis patients and experimental autoimmune prostatitis (EAP) mice. (A) CXCL10 expression in the serum of chronic prostatitis patients and healthy volunteers. (B) CXCL10 expression in the serum of chronic prostatitis patients with and without pain symptom. (C) Pearson’s correlation coefficient analysis for CXCL10 expression and National Institutes of Health Chronic Prostatitis Symptom Index. (D) The diagnostic efficiency of CXCL10 expression in the serum for chronic prostatitis patients. The expression levels of CXCL10 in prostate (E) and serum (F) of EAP mice. (G) RT-qPCR was used to assess CXCL9, CXCL10, CXCL11, and CXCR3 expressions in prostate from EAP mice. (H) IHC staining for CXCL10 and CXCR3 in prostate from EAP mice. Quantification of IHC staining of CXCL10 (I) and CXCR3 (J) in prostate from EAP mice. Data are shown as mean ± standard deviation (SD) by one-way ANOVA analysis (B) , or unpaired, two-tailed Student’s t -test analysis (A, E, F, G, I, J) , or Pearson’s correlation (C) . * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cytokine levels in mouse plasma and prostate tissue homogenate samples from immunized mice were detected using ELISA kits for CXCL9 (E-EL-M0020c, Elabscience, Wuhan, China), CXCL10 (E-EL-M0021c, Elabscience, Wuhan, China), CXCL11 (JYM1057Mo, Elisa Lab, Wuhan, China), IFN-γ (E-EL-M0048c, Elabscience, Wuhan, China), IL-6 (CSB-E04639m, Cusabio, Wuhan, China), and MCP1 (CSB-E07430m, Cusabio, Wuhan, China).

Techniques: Expressing, Diagnostic Assay, Quantitative RT-PCR, Immunohistochemistry, Standard Deviation, Two Tailed Test

Journal: Science Advances

Article Title: Versatility of bacterial outer membrane vesicles in regulating intestinal homeostasis

doi: 10.1126/sciadv.ade5079

Figure Lengend Snippet: ( A to D ) In vivo effects on repairing the gut physicochemical barrier of DSS mice after daily oral delivery of 100 μl of Akk OMV suspension containing 20 μg of total proteins or PBS for 5 days. (A) Alcian blue staining images of the proximal colon. Black and yellow arrows represent goblet cells with pathological cavities and the thickened mucus layer, respectively. Scale bars, 600 μm. (B) Percentage of goblet cells in the intestinal epithelium. (C) 3D CLSM images of the intestinal epithelial cells. Red fluorescence indicates Cy5.5-labeled Akk OMVs. Scale bars, 50 μm. (D) Immunofluorescence images of tight junctions ZO-1 and occludin expressed on the epithelial cells in the proximal colon. Scale bars, 20 μm. ( E and F ) In vitro effects on Caco-2 cells after treatment with Akk OMVs (0.1 mg/ml) along with LPS (5 μg/ml) at 37°C for 24 hours. Cy5.5-labeled OMVs were used for CLSM. (E) Immunofluorescence and (F) CLSM images. Scale bars, 25 μm. ( G ) Cytokine levels of IL-10, IL-13, and ACE2 in colon tissue together with IFN-γ and CRP in serum measured by ELISA. Samples were collected from DSS mice after treatment with Akk OMVs or PBS. Data are means ± SEM ( n ≥ 5). Significance was assessed using t test, giving P values: * P < 0.05.

Article Snippet: Levels of cytokines including IL-10 (EK210/4, MultiSciences), IL-13 (EK213/2, MultiSciences), and ACE2 (EK1188, Boster Biological Technology) in colon tissue as well as IFN-γ (EK280/3, MultiSciences) and CRP (EK294/2, MultiSciences) in serum were measured by commercially available ELISA kits.

Techniques: In Vivo, Suspension, Staining, Fluorescence, Labeling, Immunofluorescence, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: Journal of Advanced Research

Article Title: Nano-Honokiol ameliorates the cognitive deficits in TgCRND8 mice of Alzheimer’s disease via inhibiting neuropathology and modulating gut microbiota

doi: 10.1016/j.jare.2021.03.012

Figure Lengend Snippet: Nano-HO improved the spatial learning and memory functions of TgCRND8 mice. The spatial learning and memory functions were evaluated using MWMT (n = 9). (A) Experimental design and treatment schedule to evaluate the neuroprotective effects of HO and Nano-HO on TgCRND8 transgenic mice; (B) The latency to find a hidden platform during 4 consecutive days of training; (C) Time spent in the target quadrant; (D) Number of target crossings; (E) Representative swimming tracks of mice in the probe test. Nano-HO decreased the levels of cytokines including TNF-α (F), IL-1β (G) and IL-6 (H), enhanced the level of CCR2 (I) in the brain tissues of TgCRND8 mice (n = 6). Data were expressed as mean ± SEM. # p < 0.05 and ## p < 0.01 when compared with the WT group; * p < 0.05 and ** p < 0.01 when compared with the Tg + vehicle group; ▲ p < 0.05 when compared with free HO group.

Article Snippet: The levels of TNF-α (Catalog no.: ab100747, Abcam, Cambridge, UK), IL-6 (Catalog no.: ab100712, Abcam, Cambridge, UK), IL-1β (Catalog no.: ab100704, Abcam, Cambridge, UK) and CCR2 (Catalog no.: CSB-EL004841MO, CUSABIO, Houston, USA) in the supernatants were determined using commercially available ELISA kits according to the manufacturer’s instructions.

Techniques: Transgenic Assay

Journal: medRxiv

Article Title: CC- chemokine ligand 25 mediates inflammatory crosstalk between adipose and liver in non-alcoholic fatty liver disease

doi: 10.1101/2021.07.26.21261056

Figure Lengend Snippet: A serum concentration of CCL25 measured by ELISA showed a positive correlation with BMI (Pearson r 2 0.500, p<0.001). B measurement of protein levels of CCL25 measured by immunohistochemical staining showed an increase in CCL25 levels in obesity (* p<0.05 by one-way ANOVA). C Gene expression of CCL25 in adipose and liver tissue measured by rt-PCR showed greater expression in adipose tissue. D Protein levels of CCL25 were comparable between adipose and liver tissue.

Article Snippet: An ELISA kit from Cusabio (catalogue number CSB-E09189h) was used to measure concentration of CCL25 in protein lysates from whole adipose or liver tissue.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: medRxiv

Article Title: CC- chemokine ligand 25 mediates inflammatory crosstalk between adipose and liver in non-alcoholic fatty liver disease

doi: 10.1101/2021.07.26.21261056

Figure Lengend Snippet: A serum concentration of CCL25 measured by ELISA in lean and obese individuals **** p<0.001 by Mann Whitney test B lymphocyte trafficking in response to pre-treatment of hepatic endothelial cells with recombinant CCL25 ****p<0.001 by two-way ANOVA

Article Snippet: An ELISA kit from Cusabio (catalogue number CSB-E09189h) was used to measure concentration of CCL25 in protein lysates from whole adipose or liver tissue.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Recombinant

Journal: medRxiv

Article Title: CC- chemokine ligand 25 mediates inflammatory crosstalk between adipose and liver in non-alcoholic fatty liver disease

doi: 10.1101/2021.07.26.21261056

Figure Lengend Snippet: A adherence of leukocytes to HSEC after pre-treatment with CCL25 (* one-way ANOVA p<0.05) B Total CCR9 + cells isolated from normal and NAFLD liver tissue (** student’s t-test p<0.01). C CCR9 expression on leukocyte subsets from normal and NAFLD liver tissue D CCR9 expression on liver-infiltrating monocytes in normal and NAFLD liver tissue

Article Snippet: An ELISA kit from Cusabio (catalogue number CSB-E09189h) was used to measure concentration of CCL25 in protein lysates from whole adipose or liver tissue.

Techniques: Isolation, Expressing