Characterization of RNKDLS. (A) RNK2.38 or RNKDLS were stained with isotype control (black line) or anti-Ly49D (red line) and analyzed for Ly49D expression by FACS (left panels). Similar aliquots of cells were analyzed for calcium mobilization after stimulation with anti-Ly49D (4E5) and crosslinking antibody. (B) RNKD2.38 and RNKDLS were stimulated with anti-Ly49D (4E5) for the time indicated. Whole cell lysates were then probed with anti-phospho-Erk1/2 (top panel) or total Erk1/2 (bottom panel). (C) Lysates of resting RNKD2.38 or RNKDLS were immunoprecipitated with control antibody [C] or with anti-LAT and immunoblotted with anti-LAT (lanes 1-4). Whole cell lysates (WCL) from RNKD2.38 and RNKDLS were immunoblotted with anti-LAT (lanes 5,6). (D) Equal amounts of whole cell lysates of RNKDLS and RNK2.38 were immunoblotted for the indicated signaling proteins. Results are representative of at least 2 independent experiments.