American Society of Hematology

Figure 1

$Figure 1. Rituximab enhances capping of CD20 on the B-cell surface. Fluorescent and bright-field images of B-cell lines Daudi (A), 721.221 (B), and Raji (C) expressing CD20-GFP or primary B cells (D) untreated or incubated with 10 μg/mL rituximab (left and right panels, respectively). Primary B cells were additionally labeled with anti–CD20-AF647 antibody recognizing the intracellular portion of the protein. Scale bars represent 10 μm. (E) Capping of CD20 was quantified on images of Daudi/CD20-GFP and primary B cells incubated in the absence or presence of 1 μg/mL and 10 μg/mL rituximab. Graph represents mean ± SEM of 3 independent experiments. (F) Capping of CD20 in Daudi/CD20-GFP cells was quantified by flow microscopy. Cells were incubated alone or with 1 μg/mL or 10 μg/mL rituximab and then fixed and analyzed by the ImageStream multispectral imaging flow cytometer. Graph represents mean ± SEM of 2 independent experiments. (G) Quantification of the fraction of CD20 localized in the cap in Daudi/CD20-GFP and primary B cells. A total of 30 to 32 cells were analyzed per condition. Data were analyzed by 1-way ANOVA with Bonferroni post-test. ***P < .001.$

Rituximab enhances capping of CD20 on the B-cell surface. Fluorescent and bright-field images of B-cell lines Daudi (A), 721.221 (B), and Raji (C) expressing CD20-GFP or primary B cells (D) untreated or incubated with 10 μg/mL rituximab (left and right panels, respectively). Primary B cells were additionally labeled with anti–CD20-AF647 antibody recognizing the intracellular portion of the protein. Scale bars represent 10 μm. (E) Capping of CD20 was quantified on images of Daudi/CD20-GFP and primary B cells incubated in the absence or presence of 1 μg/mL and 10 μg/mL rituximab. Graph represents mean ± SEM of 3 independent experiments. (F) Capping of CD20 in Daudi/CD20-GFP cells was quantified by flow microscopy. Cells were incubated alone or with 1 μg/mL or 10 μg/mL rituximab and then fixed and analyzed by the ImageStream multispectral imaging flow cytometer. Graph represents mean ± SEM of 2 independent experiments. (G) Quantification of the fraction of CD20 localized in the cap in Daudi/CD20-GFP and primary B cells. A total of 30 to 32 cells were analyzed per condition. Data were analyzed by 1-way ANOVA with Bonferroni post-test. ***P < .001.

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