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. 2002 May 6;195(9):1207–1213. doi: 10.1084/jem.20011783

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Copyright © 2002, The Rockefeller University Press

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Figure 4. — Lower tyrosine-phosphorylation of CD22 and less SHP-1 recruitment in presence of BPC-Neu5Ac. Daudi cells were stimulated with anti-IgM in presence of Me-Neu5Ac (control) or BPC-Neu5Ac (both 250 μM). Cells were lysed and immunoprecipitated with anti-CD22 plus anti-Vav1. The anti-Vav1 I.P. served as loading control. (A) Immunoprecipitates were separated on a gel, blotted, and probed with the indicated blotting antibodies. The membrane containing the largest fragments was reprobed with anti-CD22. (B) Densitometric analysis of band intensities of the gel. Western blots were scanned and quantified. Intensities of P-tyr bands, or SHP-1 bands were divided by intensities of Vav1 bands. Vav1 was used as loading control, because the anti-CD22 antibody showed interference with the first antibody. The calculated ratio of the highest activation (3 min, Me-Neu5Ac) was set at 100%. Mean results from four experiments are shown (±SD). *P < 0.05; **P < 0.01 in Student's t test.

Figure 4. — Lower tyrosine-phosphorylation of CD22 and less SHP-1 recruitment in presence of BPC-Neu5Ac. Daudi cells were stimulated with anti-IgM in presence of Me-Neu5Ac (control) or BPC-Neu5Ac (both 250 μM). Cells were lysed and immunoprecipitated with anti-CD22 plus anti-Vav1. The anti-Vav1 I.P. served as loading control. (A) Immunoprecipitates were separated on a gel, blotted, and probed with the indicated blotting antibodies. The membrane containing the largest fragments was reprobed with anti-CD22. (B) Densitometric analysis of band intensities of the gel. Western blots were scanned and quantified. Intensities of P-tyr bands, or SHP-1 bands were divided by intensities of Vav1 bands. Vav1 was used as loading control, because the anti-CD22 antibody showed interference with the first antibody. The calculated ratio of the highest activation (3 min, Me-Neu5Ac) was set at 100%. Mean results from four experiments are shown (±SD). *P < 0.05; **P < 0.01 in Student's t test.