Journal: PLoS ONE

Article Title: Generation and Characterisation of Cisplatin-Resistant Non-Small Cell Lung Cancer Cell Lines Displaying a Stem-Like Signature

doi: 10.1371/journal.pone.0054193

Figure Lengend Snippet: Antibody staining of PT and CisR cell lines for CD133 cell surface expression was carried out by flow cytometry using a CD133/1 (AC133) phycoerythrin (PE)-labelled antibody and IgG1 isotype control antibody. The percentage CD133+ cells were plotted for all cell lines (A). Differential expression of the CSC marker CD44 was examined using an anti-human CD44 FITC-conjugated antibody and corresponding IgG2b isotype control antibody. Expression levels of CD44 were determined for all cell lines and plotted as a percentage of the tumour cell population expressing CD44 (B). Data are expressed as Mean ± SEM from three independent experiments (n = 3) Data are expressed as Mean ± SEM from three independent experiments (n = 3) ( # p<0.01, * p<0.001).

Article Snippet: Cells (1×10 6 ) were incubated with either CD133/1 (AC133) phycoerythrin (PE)-labelled antibody or isotype control antibody (IgG1) (Miltenyi Biotec GmbH), or anti-human CD44 FITC-conjugated antibody and corresponding isotype control (IgG2b) (ImmunoTools GmbH, Germany) for 30 min in the dark at 4°C.

Techniques: Staining, Expressing, Flow Cytometry, Control, Quantitative Proteomics, Marker

Journal: Cancer Immunology Research

Article Title: Tumor-Associated Neutrophils Drive B-cell Recruitment and Their Differentiation to Plasma Cells

doi: 10.1158/2326-6066.cir-20-0839

Figure Lengend Snippet: Figure 5. TANs drive B-cell differentiation independently of T cells. A, Quantification of CD138 expression on splenic B220þ cells following various culture conditions—when cultured alone (B), with T cells (B þ T, ratio 1:1), with TANs (B þ TAN, ratio 1:1), with TAN and T cells (B þ TAN þ T, ratio 1:1:1), with T cells but no contact allowed with TANs (B þT//TAN,ratio 1:1:1), or TANs cocultured with T cells but no contact allowed with B cells (B//T þTAN, ratio 1:1:1). Data represent the mean SEM (n ¼ 6–9; , P < 0.001; n.s., not significant). Groups were compared using one-way ANOVA. B, Representative flow plots displaying CD138 expression out of total B220þ population in the different coculture conditions. C, Quantification of IgG release to the media following overnight coculture of isolated splenic B220þ

Article Snippet: Isotype control antibodies were as follows: APCconjugated rat IgG2a (clone 2A3; Biogems), FITC-conjugated rat IgG2b (clone RTK4530; BioLegend), BGViolet450 rat IgG2a (clone 2A3, Biogems), PE-conjugated rat IgG2a (clone 2A3, Biogems).

Techniques: Cell Differentiation, Expressing, Cell Culture, Isolation

Journal: Cancer Immunology Research

Article Title: Tumor-Associated Neutrophils Drive B-cell Recruitment and Their Differentiation to Plasma Cells

doi: 10.1158/2326-6066.cir-20-0839

Figure Lengend Snippet: Figure 6. TANs express membranal BAFF, but not membranal APRIL, and mediate B-cell IgG production in a BAFF-R–dependent manner. A and B, Expression of membranal BAFF and APRIL in TANs within the whole tumor (A) and following TANs' isolation from the tumors (B). Representative histograms showing BAFF and APRIL expression in TANs (gated as total Ly6Gþ population) are provided (right plots). C, IgG production by splenic B cells cocultured with TANs (ratio 1:5) in the absence or presence of anti–BAFF-R antibody. Data represent the mean SEM (n ¼ 4; , P < 0.01; , P < 0.001). Groups were compared using one-way ANOVA. D, Quantification of CD138 expression on isolated splenic B220þ cells cultured alone or cocultured with TANs (ratio 1:5), without or with blocking of the three potential BAFF receptors, BAFF-R, TACI, or BCMA. Data represent the mean SEM (n ¼ 4–10; , P < 0.001; n.s., nonsignificant). Groups were compared using one-way ANOVA.

Article Snippet: Isotype control antibodies were as follows: APCconjugated rat IgG2a (clone 2A3; Biogems), FITC-conjugated rat IgG2b (clone RTK4530; BioLegend), BGViolet450 rat IgG2a (clone 2A3, Biogems), PE-conjugated rat IgG2a (clone 2A3, Biogems).

Techniques: Expressing, Isolation, Cell Culture, Blocking Assay