Journal:

Article Title: Epstein-Barr Virus Infection of Human Astrocyte Cell Lines

doi:

Figure Lengend Snippet: Analysis of CR2 expression by using MAb HB5 on wild-type and CR2-transfected astrocyte lines: T98WT (A), T98CR2 (B), CB193WT (C), CB193CR2 (D), and Raji (positive control) (E). The cells were stained with anti-CR2 MAb HB5 (open area) or with an isotype control (shaded area) and FITC-labeled goat F(ab′)2 anti-mouse IgG, as described in Materials and Methods.

Article Snippet: Monoclonal antibodies (MAbs) used for the screening and the analysis of epitopes expressed by transfected cells were HB5 (immunoglobulin G2a [IgG2a]) (ATCC; HB135) ( 52 ), purified from ascites fluid.

Techniques: Expressing, Transfection, Positive Control, Staining, Control, Labeling

Journal: Cell Death & Disease

Article Title: IL-6-C/EBPβ signaling drives monocytic differentiation of murine cultured lymphoid progenitors with immunoregulatory properties

doi: 10.1038/s41419-025-07930-4

Figure Lengend Snippet: a – c cCLPs generated from Lin - Flk2 + IL7Rα + CD27 + Ly6d - cells in murine bone marrow were maintained in cCLP medium for 3 weeks prior to the following experiments. a Representative FACS profiles of CD11b and CD115 expression 3 days after culturing 3000 cCLPs with Flt3L (FL) alone ( upper panel ) or together with IL-6 (FL/IL-6) ( lower panel ). The percentage in the gate is shown. b , c Cell number ( upper panel ) and percentage ( lower panel ) of CD11b + CD115 + cells on day 3 after culture with the indicated cytokines ( n = 3 each) or at the indicated time points ( n = 4 each) with FL ( open squares ) or FL/IL-6 ( closed squares ). d Representative FACS profiles on IL-7Rα and CD115 expression on day 7 in cCLPs cultured with the indicated cytokines. The percentages in the gate are shown. e Percentage of CD115 + IL-7Rα - cells ( n = 3 each) in ( d ). f Histograms of IL-6Rα ( upper panel ) and gp130 ( lower panel ) expression ( red histograms ) on cCLPs, with isotype control shown in gray . Mean fluorescence intensity (MFI) values are shown in each panel. g pSTAT3 expression in cCLPs at 0 ( gray ) or 15 min ( red ) after the addition of IL-6 (100 ng/ml). The dotted line represents the isotype control. h May-Grünwald Giemsa staining images of cCLPs (control, left panel ) or CD11b + CD115 + Ly6C + CCR2 + cells isolated by FACS 4 days after cCLP culture with FL/IL-6/SCF ( middle panels ). The right panels show a representative image of CD11b + CD115 + Ly6G - Ly6C + CCR2 + monocytes isolated ex vivo (BMmono) or cultured macrophages (BMDMs) from the bone marrow of C57BL/6 mice. Scale bar = 10 μm. i cCLPs were cultured with 10 ng/mL of FL/IL-6/SCF for 4 days. A representative FACS profile of indicated molecules. The percentages in the gate and histograms are shown. Data are mean ± SD with statistical significance determined by one-way ANOVA (in b , e ) or two-way ANOVA (in c ). The p-values are represented as **, <0.01; ***, <0.001; ****, <0.0001. n.s., not significant. All data are representative of at least two independent experiments.

Article Snippet: Single-cell suspensions were prepared in FACS buffer PBS containing 1% FBS, 5 mM EDTA, and 0.2% sodium azide and blocked with anti-CD16/CD32 (clone 2.4G2) before staining with the following fluorochrome-conjugated anti-mouse monoclonal antibodies: FITC-labeled Ly6G (1A8, #127605), Ly6C (1G7.G10, #128006), and I-A/I-E (M5/114.15.2, #107605) from BioLegend, and PD-L1 (MIH6, MCA2626F) from AbD Serotec; PE-labeled IL-6Rα (W18166A, #160405), Flk2 (#135305), Ly6d (49-H4, #138606), I-A/I-E (M5/114.15.2, #107607), and B220 (RA3-6B2, #103207) from BioLegend; PE/Cy7-labeled CD11b (M1/70, #101215), Ly6C (1G7.G10, #128018), and IL-7Rα (A7R34, #135014) from BioLegend; APC-labeled CD115 (AFS98, #135509), gp130 (4H1B35, #149405), F4/80 (BM8, #123116), CD27 (LG.3A10, #124211), and PD-L2 (TY25, #107206) from BioLegend; Alexa 700-labeled Sca1 (D7, #108141) from BioLegend; APC/Cy7-labeled Ly6G (1A8, #127624), Ly6C (1G7.G10, #128025), I-A/I-E (M5/114.15.2, #107627), and B220 (RA3-6B2, #103224) from BioLegend; APCVio770-labeled Ly6d (49-H4, #130-115-315) from Miltenyi Biotec (Germany); BV421-labeled CD115 (AFS98, #135513), F4/80 (BM8, #123131), and c-Kit (2B8, #105827) from BioLegend; BV605-labeled CD19 (1D3, #115539) from BioLegend; BV650-labeled CD11b (M1/70, #101239) from BioLegend; PerCP/Cy5.5-labeled Streptavidin (#405214) from BioLegend; and PE-labeled CCR2, either Y15-488 from BD Biosciences (San Jose, CA, USA, #475301) or clone 475301 from R&D Systems (Minneapolis, MN, USA,FAB5538P).

Techniques: Generated, Expressing, Cell Culture, Control, Fluorescence, Staining, Isolation, Ex Vivo