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Figure 2 | BMC Immunology

Figure 2

From: Human CD72 splicing isoform responsible for resistance to systemic lupus erythematosus regulates serum immunoglobulin level and is localized in endoplasmic reticulum

Figure 2

CD72fl but not CD72Δex8 is efficiently expressed on B cell surface and regulates BCR signaling. A: Cell surface expression of NP-reactive BCR and human CD72 in K46μv transfectants. Cell surface expression was analyzed by flow cytometry using a FACSCalibur. B: ERK phosphorylation induced by antigen NP15-coupled BSA (NP-BSA) stimulation in K46μv transfectants. Cells were stimulated with 0.2 μg/ml NP-BSA. Cells were lysed, and total cell lysates were subjected to Western blot analysis. The same blots were reprobed with anti-β-tubulin Ab to ensure equal loading. C: Ca2+ influx induced by NP-BSA stimulation in K46μv transfectants. Transfectants were loaded with fluo-4-AM, and intracellular free Ca2+ was measured with a FACSCalibur. Transfectants were stimulated with 0.2 μg/ml NP-BSA at 30s (indicated by arrow), and measurement of free Ca2+ was continued for 180 s. D: Phosphorylation and SHP-1 recruitment of CD72 induced by NP-BSA stimulation in K46μv transfectants. Cells were lysed, and CD72 was immunoprecipitated with anti-FLAG Ab. Immunoprecipitates (IP) were subjected to Western blot analysis. The same blots were reprobed with anti-CD72 Ab to ensure equal loading. E: Expression of CD72 in total cell lysates in K46μv transfectants. Cells were lysed, and total cell lysates were subjected to Western blot analysis. The same blots were reprobed with β-tubulin Ab to ensure equal loading. Representative data from three experiments are shown.

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