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: Antioxidants (Basel, Switzerland)

Article Title: Aster-B Modulates Oxidative Stress Responses and Carotenoid Distribution in ARPE-19 Cells.

doi: 10.3390/antiox14050575

Figure Lengend Snippet: Figure 1. CRISPR/dCas9 activation of endogenous GRAMD1B gene expression in ARPE19 GRAMD1B-null cells. (A) ARPE-19 cells were transfected with scrambled DNA at 0.5, 1, and 1.5 µg DNA per well and GRAMD1B plasmid DNA at 0.5, 1, and 1.5 µg DNA per well, respectively. Western blot analysis was conducted with 50 µg of total protein per lane and showed that GRAMD1B is expressed before clonal selection and becomes enriched in post-clonal ARPE-19 cells (25 and 50 µg per well) treated with GRAMD1B plasmid DNA (1.5 µg per well). In contrast, GRAMD1B expression is absent in control cells treated with scrambled control DNA (1.5 µg per well). A549 cells were used as a positive control for GRAMD1B expression. β-Actin was used as a loading control. (B) Confocal imaging analysis with ARPE-19 cells transfected with plasmid DNA confirmed that GRAMD1B protein is expressed and mostly localized in the cytoplasm. Similar results were obtained from WB and ICC in more than three independent experiments. The scale bar is 10 µm.

Article Snippet: The antibodies used included GRAMD1B (Proteintech), COXIV, Caspase-3, PARP, and Histone H3 (Cell Signaling Technology) at a dilution of 1:1000.

Techniques: CRISPR, Activation Assay, Gene Expression, Transfection, Plasmid Preparation, Western Blot, Selection, Expressing, Control, Positive Control, Imaging