Journal: Research

Article Title: Effect of Microcurrent Stimulation on Cartilage Regeneration in Engineered Chondrocyte Spheroids

doi: 10.34133/research.1462

Figure Lengend Snippet: Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) Representative Live/Dead staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.

Article Snippet: Cell metabolic activity was measured using the CCK-8 kit (Beyotime Biotechnology, Shanghai, China).

Techniques: In Vitro, Cell Culture, Control, CCK-8 Assay, Staining, Analysis

Journal: Inflammation Research

Article Title: IL-1β/IL-6 inflammatory crosstalk drives DNA methylation and histone remodeling associated with cytokine induced senescence in luminal breast cancer

doi: 10.1007/s00011-026-02391-7

Figure Lengend Snippet: a Quantification of percentage of cells in G 0/ G 1 , S, and G2/M cell cycle phases under control, IL-1β, IL-1β + IL-1Ra, and IL-1Ra treatment conditions for 24 h and 72 h treatments in MCF-7 cells through flow cytometry. b CCK-8 assay results for control, IL-1β, IL-1β + IL-1Ra and IL-1Ra treatment conditions for 24 h and 72 h treatments in MCF-7 cells. c EdU incorporation assay in MCF-7 cells to measure DNA synthesis for control, IL-1β, IL-1β + IL-1Ra and IL-1Ra treatment conditions for 24 h and 72 h in MCF-7 cells. For statistical significance assessment Two-way ANOVA with Tukey’s multiple comparisons test was used for cell cycle analysis and ordinary one-way ANOVA with Tukey’s multiple comparisons test was used for CCK8 and EdU assays (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). d Quantification of apoptotic stages post 24 h and 72 h treatments in MCF-7 cells for control, IL-1β, IL-1β + IL-1Ra and IL-1Ra treatment conditions. e Quantification of intracellular ROS measured as DCFDA mean fluorescence intensity (MFI) post IL-1β treatment for 24 and 72 h. One-way ANOVA with Tukey’s multiple comparisons test was used for significance testing (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). f Representative images of SA-β-gal staining for control, IL-1β, IL-1β + IL-1Ra, and IL-1Ra treated MCF-7 cells post treatment for 72 h and quantification of senescence-associated β-galactosidase (SA-β-gal) positivity. At least 100 cells per field were manually counted across ten random fields (10×), and the percentage of SA-β-gal–positive cells was calculated as the mean across fields. Statistical significance was assessed using ordinary one-way ANOVA with Tukey’s multiple comparisons test (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). g Flow cytometric analysis of cellular senescence using the CellEvent™ Senescence Green assay. Median fluorescence intensity (X-Med) of the gated Senescence Green-positive population in control, IL-1β, IL-1β + IL-1Ra, and IL-1Ra-treated MCF-7 cells following 72 h of treatment was used for measuring senescence. One-way ANOVA with Tukey’s multiple comparisons test was used for significance testing (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: Cell counting kit-8 (CCK-8) assay (Dojindo, Japan) was used to assess proliferative effects of cytokine induction on MCF-7 breast cancer cells.

Techniques: Control, Flow Cytometry, CCK-8 Assay, EdU Assay, DNA Synthesis, Cell Cycle Assay, Fluorescence, Staining, Analysis

Journal: Inflammation Research

Article Title: IL-1β/IL-6 inflammatory crosstalk drives DNA methylation and histone remodeling associated with cytokine induced senescence in luminal breast cancer

doi: 10.1007/s00011-026-02391-7

Figure Lengend Snippet: a CCK-8 and b EdU incorporation assays demonstrating increased proliferation and DNA synthesis following IL-1β conditioned media (CM) treatment, with partial reversal by IL-1Ra or IL-6R blockade. Ordinary one-way ANOVA with Tukey’s multiple comparisons test was used to assess significance (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001). c Quantification of percentage of cells in G 0/ G 1 , S, and G2/M cell cycle phases in naïve recipient MCF-7 cells following 24-hour treatment with conditioned media from the indicated treatment groups, including IL-1β-derived conditioned media with or without huE17 through flow cytometry. Two-way ANOVA with Tukey’s multiple comparisons test was used for cell cycle analysis to determine statistical significance. d Soft agar colony formation assay to assess anchorage-independent growth under IL-1β CM, attenuated by IL-1Ra or antibody treatment conditions and quantification of mean colony diameter. One-way ANOVA with Dunnett’s test was used for significance (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001)

Article Snippet: Cell counting kit-8 (CCK-8) assay (Dojindo, Japan) was used to assess proliferative effects of cytokine induction on MCF-7 breast cancer cells.

Techniques: CCK-8 Assay, EdU Assay, DNA Synthesis, Derivative Assay, Flow Cytometry, Cell Cycle Assay, Soft Agar Assay, Colony Assay, Tube Formation Assay