Journal: Frontiers in Cell and Developmental Biology

Article Title: Amlexanox and Forskolin Prevents Isoproterenol-Induced Cardiomyopathy by Subduing Cardiomyocyte Hypertrophy and Maladaptive Inflammatory Responses

doi: 10.3389/fcell.2021.719351

Figure Lengend Snippet: ALX and FSK combination preserves mice myocardial architecture during CCS. (A) Representative whole hearts from all experimental groups. (B) Representative Masson’s trichrome stained longitudinal section of whole hearts from all groups. (C) Representative Masson’s trichrome stained transverse section of whole hearts from all groups. (D–G) Representative Western blots and graphical presentations cleaved caspase 3, collagen I, and collagen III compared between Vhl and PCH mice, and ALX treatment, FSK treatment, and ALX and FSK combination treatment ( n = 4 hearts per treatment group). Western blots were performed in triplicates, and each protein band in the representative blot is an independent biological sample. (H,I) Representative microscopic images of Masson’s trichrome staining and collagen volume fraction (CVF) in the ventricular tissue sections from all groups. The plotted values are the means of the CVF from each mouse ( n = 6–8 fields of view per 5–7 sections per 8–16 hearts per group). (J,K) Representative microscopic images of wheat germ agglutinin (WGA) merged with DAPI staining and graphical presentation of measured cardiomyocyte diameters from ventricular tissue sectionings across all groups. The plotted values are the means of the cardiomyocyte sizes from each mouse ( n = 10–12 cells per five fields of view per five sections per six to seven hearts per group). Representative cardiomyocytes are marked in yellow boxes, and their zoomed-in (× 5) inserts to show hypertrophy are marked in red boxes. The Vhl mice group is representative of the Ctrl mice group in results illustrations due to similarity in their data. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 among the therapeutic groups; &&& p < 0.001 vs. Vhl; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. the therapeutic groups. CVF was estimated by dividing the collagen area with the total myocardial area and multiple by 100. Data are expressed as mean ± SEM. Data were analyzed using one-way ANOVA, followed by Tukey’s post hoc analysis.

Article Snippet: PCH preventive models were made by intraperitoneal injection of ALX (ab142825; Abcam) 2.5 mg/100 g/day and/or FSK (1099; Tocris Bioscience, United Kingdom) 0.5 mg/100 g/day.

Techniques: Staining, Western Blot

Journal: Frontiers in Cell and Developmental Biology

Article Title: Amlexanox and Forskolin Prevents Isoproterenol-Induced Cardiomyopathy by Subduing Cardiomyocyte Hypertrophy and Maladaptive Inflammatory Responses

doi: 10.3389/fcell.2021.719351

Figure Lengend Snippet: ALX and FSK combination prevents proinflammatory responses aggravation in myocardia during CCS by preventing necrosis. (A) Graphical presentations of evaluated sera concentrations of troponin I from groups. The sera troponin I analysis was performed in triplicates ( n = 8 mice per treatment group). ∗ p < 0.05, ∗∗∗ p < 0.001, among the therapeutic groups; &&& p < 0.001 vs. ISO (PCH); ## p < 0.01, ### p < 0.001, vs. the therapeutic groups. (B,C) Representative CD68 IHC staining and graphical presentation of the extent of inflammatory cells infiltration into the myocardial of all groups. The Vhl mice group is representative of the Ctrl mice group in results illustrations due to similarity in their data ( n = 6–8 field of view per six to eight sections per six to seven hearts per group). (D–H) Graphical presentations of inflammatory cytokines (IL-1β, IL-6, TNFα, IL-10, and TGF-β) were evaluated by ELISA from all groups. The cytokines analysis was performed in triplicates ( n = 7–8 mice per treatment group). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, among the therapeutic groups; &&& p < 0.001 vs. Vhl; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. the therapeutic groups. Data are expressed as mean ± SEM. Data were analyzed using one-way ANOVA, followed by Tukey’s post hoc analysis.

Article Snippet: PCH preventive models were made by intraperitoneal injection of ALX (ab142825; Abcam) 2.5 mg/100 g/day and/or FSK (1099; Tocris Bioscience, United Kingdom) 0.5 mg/100 g/day.

Techniques: Immunohistochemistry, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Altered Intestinal Morphology and Microbiota Composition in the Autism Spectrum Disorders Associated SHANK3 Mouse Model

doi: 10.3390/ijms20092134

Figure Lengend Snippet: Altered gut morphology in Shank3αβ knock-out (KO) mice. ( A – D ) Histological evaluation of GI tract from wild type and Shank3αβ KO mice. ( A ) Longitudinal cross sections of Shank3αβ KO mice and wild type (WT) mice were stained with hematoxylin/eosin (HE) (upper panels) and periodic acid schiff (PAS) reaction (lower panels). Exemplary images are shown. ( B – D ) Morphological analysis of ( B ) villi length and ( C ) width, and ( D ) crypt depth reveals a significantly decreased villi length (Mann-Whitney U -test, p = 0.009; n = 5 animals per group) but not width ( p = 0.534), and normal crypt depth ( p = 0.983) in Shank3αβ KO mice. ( E , F ) Immunohistochemistry was performed on 5 mice per group and 5 optic fields of view each from 3 sections per mouse were analyzed. ( E ) A slight but non-significant decrease in FABP2 signal intensity was observed in Shank3αβ KO mice compared to wild types (left panel). Significantly higher ZONULIN-1 levels were found in Shank3αβ KO mice (right panel) ( t -test, p = 0.0413). ( F ) The levels of CLAUDIN3 and lipopolysaccharide (LPS) were not significantly different between Shank3αβ KO mice and wild types in gut epithelium. ( G ) Significantly higher ZONULIN-1 levels in Shank3αβ KO mice were confirmed by western blotting using gut epithelium protein lysate ( t -test, p = 0.0434, n = 3 per group). ( H ) Protein lysate from liver tissue from WT and Shank3αβ KO mice ( n = 3 per group) were analyzed for E. coli LPS levels using Western Blotting. The results show significantly higher LPS levels in the liver of Shank3αβ β KO mice ( t -test, p = 0.0452). * p < 0.05, ** p < 0.01.

Article Snippet: Zonulin 1 antibody was purchased from Thermo Fisher Scientific (Invitrogen) (Waltham, MA, USA); Claudin3 antibody from Abcam (Berlin, Germany); FABP2 antibody from Thermo Fisher Scientific (Invitrogen); LPS antibody from Origene (Rockville, MD, USA); and IL6 antibody was purchased from Cell signaling Technologies (Danvers, MA, USA); GFAP antibody was purchased from Sigma Aldrich (St. Louis, MO, USA); Cytokeratin and Vimentin antibody from Abcam.

Techniques: Knock-Out, Staining, MANN-WHITNEY, Immunohistochemistry, Western Blot

Journal: mSystems

Article Title: Association of intratumoral microbiome diversity with hepatocellular carcinoma prognosis

doi: 10.1128/msystems.00765-24

Figure Lengend Snippet: Bacterial LPS and LTA are present inside HCC tissues. (a) Representative images of multiplex immunofluorescence (multiplex IF, 20×) staining with an Opal kit. The bacteria were stained with antibodies against LPS and LTA, while CD45 and CD68 were used to identify immune cells. The area circled by the green curve indicates the IRR. Scale bars, 50 µm. (b) The boxplot of bacterial LPS and LTA intensity. Nine ROIs were selected.

Article Snippet: Staining was conducted manually using primary antibodies against the following markers: CD45 (anti-CD45, eBioscience #14-0459-82), CD68 (anti-CD68, Invitrogen #MA5-12407), LPS (anti-LPS, abcam #ab35654), and LTA (anti-LTA, Santa Cruz #sc57752).

Techniques: Multiplex Assay, Immunofluorescence, Staining, Bacteria