Journal: iScience

Article Title: Molecular maps of synovial cells in inflammatory arthritis using an optimized synovial tissue dissociation protocol

doi: 10.1016/j.isci.2024.109707

Figure Lengend Snippet:

Article Snippet: MACS BSA Stock Solution , Miltenyi Biotec , Cat# 130-091-376.

Techniques: Recombinant, Red Blood Cell Lysis, Staining, Control, Blocking Assay, Multiplex Assay, Software, Transferring

Journal: Heliyon

Article Title: Inhibition of CTLA-4 accelerates atherosclerosis in hyperlipidemic mice by modulating the Th1/Th2 balance via the NF-κB signaling pathway

doi: 10.1016/j.heliyon.2024.e37278

Figure Lengend Snippet: The anti-CTLA-4 antibody promotes CD4 + T-cell infiltration and T-cell differentiation into Th1 cells in the aortic root. (A) GO enrichment analysis showing the significantly enriched biological processes in hyperlipidemic mice administered the anti-IgG antibody or anti-CTLA-4 antibody. n = 3 per group. (B) GSEA of RNA-seq data showing the enrichment of “regulation of Th1-type immune response” and “Th1 cell differentiation” in hyperlipidemic mice administered the anti-IgG antibody or anti-CTLA-4 antibody. NES: normalized enrichment score; FDR: adjusted p value. (C) Representative immunofluorescence staining of CD4 (red), α-SMA (green), and DAPI (blue) in mice in the indicated groups. The square represents the colocalization of CD4 with the smooth muscle marker α-SMA. The scale bars are 100 μm and 20 μm. (D) Quantitative analysis of CD4 fluorescence intensity in aortic roots in the indicated groups. The data are presented as the means ± SEM (n = 6). (E) Flow cytometric analysis of the distributions of CD4 + IFN-γ + T cells and CD4 + IL-4 + T cells in peripheral blood in the indicated groups. (F) Relative quantification of the percentages of CD4 + IFN-γ + T cells and CD4 + IL-4 + T cells among the sorted CD4 + T cells. The data are presented as the means ± SEM (n = 5). (G) The Th1/Th2 ratio was estimated by determining the ratio of CD4 + IFN-γ + IL-4 − cells to CD4 + IFN-γ − IL-4 + cells. The data are presented as the means ± SEM (n = 5). (H) Quantitative results were obtained by real-time PCR showing the relative mRNA expression of IFN-γ and IL-4 in the indicated groups. The data are presented as the means ± SEM (n = 6). *P < 0.05 vs . the anti-IgG group. (I – J) Quantitative analyses demonstrating the number of CD4 + IFN-γ + T cells and CD4 + IL-4 + T cells in aortic roots from each indicated experimental group. The data are presented as the means ± SEM (n = 4). *P < 0.05 vs . the anti-IgG group.

Article Snippet: One hundred microliters of mouse peripheral blood were collected in isotonic RPMI 1640 complete culture medium (R8758, Sigma-Aldrich) and stimulated with 2 μL of leukocyte activation cocktail (550583, BD Biosciences) in 5 % CO 2 at 37 °C for 4 h. Then, the cells were treated with anti-mouse CD3 (APC-Cy7-CD3, 1:200, 557596, BD Biosciences) and anti-mouse CD4 (BV510-CD4, 1:200, 563106, BD Biosciences) for 30 min and lysed in red blood cell lysis solution (130-094-183, Miltenyi Biotec) for 10 min. After being treated with fixation/permeabilization concentrate at room temperature for 1 h, total T lymphocytes were further stained with anti-mouse IFN-γ (FITC-IFN-γ, 1:100, 554411, BD Biosciences) and anti-mouse IL-4 (PE-IL-4, 1:100, 554435, BD Biosciences) for 30 min. Isotype controls were used for compensation and to confirm antibody specificity.

Techniques: Cell Differentiation, RNA Sequencing, Immunofluorescence, Staining, Marker, Fluorescence, Quantitative Proteomics, Real-time Polymerase Chain Reaction, Expressing

Journal: International Journal of Molecular Sciences

Article Title: New 2-[(4-Amino-6- N -substituted-1,3,5-triazin-2-yl)methylthio]- N -(imidazolidin-2-ylidene)-4-chloro-5-methylbenzenesulfonamide Derivatives, Design, Synthesis and Anticancer Evaluation

doi: 10.3390/ijms23137178

Figure Lengend Snippet: Induction of apoptosis by compounds 48 and 140 in MCF-7 cells. Cells were treated with the indicated concentrations of compounds 48 and 140 for 72 h, stained with Annexin V-PE and 7-AAD and analyzed with flow cytometry. ( A ) Dot plots showing early apoptotic (bottom right quadrant), late apoptotic (upper right quadrant), viable (lower left quadrant) and necrotic cell populations (upper left quadrant). ( B ) Graphs indicating early and late apoptotic cells. Values represent the mean ± SD of three repetitions. Data were analyzed by one-way ANOVA with Tukey’s post hoc test [ p < 0.05 (*), p < 0.01 (**), p < 0.001 (***)].

Article Snippet: After staining, cells were diluted in Annexin V binding buffer and analyzed on a BD FACSCalibur flow cytometer (BD Biosciences).

Techniques: Staining, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: New 2-[(4-Amino-6- N -substituted-1,3,5-triazin-2-yl)methylthio]- N -(imidazolidin-2-ylidene)-4-chloro-5-methylbenzenesulfonamide Derivatives, Design, Synthesis and Anticancer Evaluation

doi: 10.3390/ijms23137178

Figure Lengend Snippet: Induction of apoptosis by compounds 48 and 140 in T47D cells. Cells were treated with the indicated concentrations of compounds 48 and 140 for 72 h, stained with Annexin V-PE and 7-AAD, and analyzed with flow cytometry. ( A ) Dot plots showing early apoptotic (bottom right quadrant), late apoptotic (upper right quadrant), viable (lower left quadrant) and necrotic cell populations (upper left quadrant). ( B ) Graphs indicating early and late apoptotic cells. Values represent the mean ± SD of three repetitions. Data were analyzed by one-way ANOVA with Tukey’s post hoc test [ p < 0.01 (**), p < 0.001 (***)].

Article Snippet: After staining, cells were diluted in Annexin V binding buffer and analyzed on a BD FACSCalibur flow cytometer (BD Biosciences).

Techniques: Staining, Flow Cytometry