Figure 1. Crk and Crkl are deleted in T cells of CRK/CRKL Dko mice.

(A) CD4⁺ T cells were purified from lymph nodes of CRK/CRKL Dko and WT mice. Preactivated T cells were prepared by stimulating with plate-bound anti-CD3 and anti-CD28 for 2 days and culturing without stimuli for an additional 5 days. Whole cell lysates were analyzed by SDS-PAGE and immunoblotted with anti-CRKL, anti-CRK, and anti-ZAP70. (B) (C) Preactivated CD4⁺ T cells made as in A were fixed and permeabilized, and the intracellular staining of CRK (B) and CRKL (C) was determined by flow cytometry. (D) Thymus, spleens, and lymph nodes (LNs) were isolated from WT and CRK/CRKL Dko mice. Single-cell suspensions were analyzed by flow cytometry using the indicated antibodies. (E) Gated CD4⁺ and CD8⁺ T cells from lymph nodes (mixed peripheral and mesenteric lymph nodes) were analyzed for the indicated surface markers.