Skip to main content
. Author manuscript; available in PMC: 2016 Nov 1.
Published in final edited form as: J Immunol. 2015 Sep 28;195(9):4218–4227. doi: 10.4049/jimmunol.1501200

FIGURE 1.

FIGURE 1

Tyrosine phosphorylation of Cbl-b requires CD28 costimulation in primary T cells. (A) B6 T cells were stimulated with anti-CD3 or anti-CD3 plus anti-CD28, or treated with pervanadate, and lysed in RIPA buffer containing 1% SDS. The cell lysates were immunoprecipitated with anti-Cbl-b, and blotted with anti-pTyr and anti-ubiquitin, respectively, or blotted with phospho-specific Abs against Lck (Y394), TCRζ (Y142) ZAP-70 (Y319), LAT (Y132), and SLP-76 (Y113). Aliquots of cell lysates from each sample were blotted with anti-actin as a loading control. (B) Jurkat T cells were stimulated with anti-CD3 or anti-CD3 and anti-CD28, and lysed. The cell lysates were immunoprecipitated with anti-anti-Cbl-b, and blotted with anti-pTyr. Aliquots of cell lysates from each sample were blotted with anti-actin as a loading control. The fold changes of protein bands indicated in arbitrary densitometric units were determined by the ImageJ 1.48. Data are representative of at least four independent experiments.