Journal: Journal of Cellular and Molecular Medicine

Article Title: Regulation of Autophagy and Metabolism in Hepatocellular Carcinoma: Involvement of Wnt‐β‐Catenin Pathway

doi: 10.1111/jcmm.71070

Figure Lengend Snippet: Inhibition of cell viability and colony formation, and induction of apoptosis by riluzole. (A) Expression of stem cell markers in HCC CSCs. CSCs were stained with CD44 and CD133 antibody and the expression of CD44 and CD133 was measured by immunocytochemistry. (B, C) HCC cell lines (HepG2 and Hep3B), cancer stem cells (CSCs), and human normal hepatocytes were seeded in 96‐well plates, treated with or without riluzole (0–40 μM) for 72 h, and cell viability was measured by CellTiter‐Glo Luminescent Cell Viability Assay (Promega). (D) Colony formation. HCC cell lines (HepG2, Hep3B, SNU‐382 and SNU‐475) and CSCs were treated with riluzole (0–20 μM). Number of colonies formed at 21 days was counted under a microscope. (E) Normal human hepatocytes, HCC cell lines (HepG2 and Hep3B) and CSCs were treated with riluzole (0–20 μM) for 72 h. Apoptosis was measured by TUNEL assay. Data represent mean ± SD ( n = 4). *, # and % = significantly different from control and each other; p < 0.05.

Article Snippet: Human normal hepatocytes were purchased from ATCC.

Techniques: Inhibition, Expressing, Staining, Immunocytochemistry, Cell Viability Assay, Microscopy, TUNEL Assay, Control

Journal: Nature Communications

Article Title: Loss of Fsr quorum sensing promotes biofilm formation and worsens outcomes in enterococcal infective endocarditis

doi: 10.1038/s41467-026-68366-8

Figure Lengend Snippet: A Z-projections of WT and Δ gelE vegetations at 72 hpi, captured with LSCM and stained for DNA, myeloperoxidase (MPO), and IL-1β. Cyan insets are highlighting the presence of IL-1β between the NETs-biofilm interface. Orange inset is shown in Fig. S4B, highlighting the colocalization of IL-1β with neutrophils. Representative images shown from n = 3, N = 1. Bf = biofilm, scale = 20 µm. B Detection and quantification of IL-1β in WT and Δ gelE vegetations at 72 hpi with western blotting. Loading control = β-actin. Mean ± SEM, n = 5 from N = 2, A.U. = arbitrary units. Statistical significance was assessed with a two-tailed t-test. C Rat pro-IL-1β and its cleaved fragments in supernatants from OG1RF WT and mutant strains detected by western blotting. Rat pro-IL-1β was incubated in BHI with indicated OG1RF strains for 2, 4, 6 h. BHI = Negative control with only media and pro-IL 1β. Representative blot shown from N = 2. D Human pro-IL-1β and its cleaved fragments in supernatants from OG1RF WT and mutant strains detected by western blotting. Human pro-IL-1β was incubated in BHI with indicated OG1RF strains for 2, 4, 6, and 24 h. Gelatinase presence was also determined in these supernatants. Δ gelE :: gelE E352A expresses proteolytically inactive gelatinase. Representative blot shown from N = 2. BHI = Negative control with only media and pro-IL 1β. E Mean activation of HEK-Blue IL-1R reporter cells by supernatants harvested from OG1RF WT and Δ gelE :: gelE E352A cultures with or without human pro-IL-1β at 18 h. Stimulation of cells with mature human IL-1β was used as a positive control. N = 3, Error = SEM. Statistical significance was determined with one-way ANOVA. F Schematic representation of gelatinase (blue) and caspase-1 cleavage sites (red) across human, rat, and mouse pro-IL-1β; n = animals per group, N = independent experiments, ns not significant (p ≥ 0.05), Red arrowhead = pro-IL-1β, blue arrowhead = mature IL-1β. Exact p values are reported in the figure. Source data are provided as a Source Data file.

Article Snippet: E. faecalis overnight cultures of OG1RF WT and mutants were diluted 1:10 in 200 μl fresh BHI supplemented with human pro-IL-1β (100 ng/mL; Cat. Nr. 10139-H07E, Sino Biological) and incubated for 2, 4, 6 and 24 h, or with rat pro-IL-1β (10 ng/ml, Cat. Nr. 80023-R07E, Sino Biological) and incubated at 2, 4, and 6 h at 37 °C, static conditions.

Techniques: Staining, Western Blot, Control, Two Tailed Test, Mutagenesis, Incubation, Negative Control, Activation Assay, Positive Control