Journal: Frontiers in Immunology

Article Title: Immunomodulation by glucocorticoid-induced leucine zipper in macrophages: enhanced phagocytosis, protection from pyroptosis, and altered mitochondrial function

doi: 10.3389/fimmu.2024.1396827

Figure Lengend Snippet: Protection from Pyroptosis in GILZ-Overexpressing Macrophages. (A) Volcano plot of all differentially expressed genes (DEGs, p < 0.05, shown in grey) from untreated BMMs comparing GILZ KO and GILZ TG. Shown in black are all genes annotated in the GO term ‘cell death’ (GO:0008219), from which those with a fold change < 0.67 or > 1.5 are shown in red and counted, corresponding to the indicated n number. Normalized green (Caspase-3/7 activity) (B) and red (Cytotoxicity) (C) signal of BMMs upon ATP treatment measured by live cell imaging with the Incucyte ® S3 system of BMMs pre-treated with LPS (n = 6 mice per group). (D) IL-1β secretion measured by ELISA of ATP/LPS-treated BMMs (n = 4 mice per group). (E) Representative brightfield and red fluorescence channel images and (F) DHE signal from live cell imaging with the Incucyte ® S3 system in untreated BMMs (co) and 30 min after the addition of ATP on LPS-treated BMMs with cell culture media containing DHE relative to stimulated WT (set to 100%, n = 4–9 mice per group). Scale bar: 700 µm. (G) MitoSOX™ signal from live cell imaging with the Incucyte ® S3 system in untreated BMMs (co), 30 min after the addition of either ATP on LPS-treated BMMs (ATP+LPS) or Antimycin A (AA) relative to WT (set to 100%). p values were generated from one-way ANOVA comparison between groups for single time points and two-way ANOVA comparison for multiple time points both followed by Bonferroni post-hoc tests. *** p < 0.0001.

Article Snippet: RPMI-1640 cell culture media (#R0833), 200 mM L-glutamine (#G7513), 1% penicillin/streptomycin (#P4333), Accutase (#A6964), mitomycin C (#10107409001), antimycin A from Streptomyces sp. (#A8674), gentamicin (#G1397), dihydroethidium (DHE) (#37291), and gelatin type A (#G1890) were from Sigma-Aldrich.

Techniques: Activity Assay, Live Cell Imaging, Enzyme-linked Immunosorbent Assay, Fluorescence, Cell Culture, Generated, Comparison

Journal: Current Issues in Molecular Biology

Article Title: Beeswax Alcohol Prevents Low-Density Lipoprotein Oxidation and Demonstrates Antioxidant Activities in Zebrafish Embryos and Human Subjects: A Clinical Study

doi: 10.3390/cimb46010026

Figure Lengend Snippet: A comparative effect of beeswax alcohol (BWA) and coenzyme Q 10 (CoQ 10 ) to prevent carboxymethyllysine (CML)-induced toxicity in zebrafish embryos. ( A ) Survivability rate of zebrafish embryos at 5 h post-treatment. ( B ) Images representing developmental deformities at 5 h and 24 h post-treatment. The red and black arrows represent the embryo death and tail curvature, respectively, while the green and blue arrows represent pericardial and yolk sac edema, respectively. ( C ) Fluorescent images of dihydroethidium (DHE) and acridine orange (AO) representing the reactive oxygen species (ROS) production and apoptosis in the embryos of zebrafish. ( D ) Quantification of the fluorescent intensity. Image J software was utilized to compute the fluorescent intensity of DHE (red fluorescence) and AO (green fluorescence) staining. PBS and CML groups represent the embryos microinjected with 10 nL of PBS and 500 ng CML/10 nL PBS, respectively, while embryos in CML + BWA or CoQ 10 groups were microinjected with 500 ng CML suspended in 10 nL of BWA or CoQ 10 (final 15 μM). ** represents p < 0.01, * represents p < 0.05 compared with the AO fluorescent intensity of only-CML-injected group, while ## represents p < 0.01, and # represents p < 0.05 compared with the DHE fluorescent intensity of only-CML-injected group; “ns” represents a nonsignificant difference between the groups.

Article Snippet: 2,2-Diphenyl-1-picrylhydrazyl (Cat. No. 044150), paraoxon-ethyl (Cat. No. 36186), N -ε-carboxymethyllysine (Cat No. 14580-5 g), coenzyme Q 10 (CoQ 10 ) (Cat. No. C9538-100 mg), dihydroethidium (Cat. No. 37291), and acridine orange (Cat. No. A9231) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Software, Fluorescence, Staining, Injection