Expression and function of signaling adaptors. (A) NKG2D (top) and Ly49D (bottom) were immunoprecipitated from RMA and RMA-H60-exposed NK cells. Immunoprecipitates (IP) were analyzed using DAP-10 or KARAP/DAP-12 Abs. The results shown are representative of 2 independent experiments performed. (B) Total cellular mRNA was extracted from RMA and RMA-H60-exposed NK cells. The relative abundance of specific transcripts was estimated by RT-PCR analysis on serial 10-fold cDNA dilutions. The results shown are representative of 5 independent experiments performed. (C) Immunoblot analysis of total cell lysates of day-3 cocultures using the indicated Abs. The results shown are representative of 4 independent experiments performed. (D) KARAPki NK cells were cocultured with RMA (○) or RMA-H60 (•) for 3 days. The lytic activity of KARAPki NK cells toward RMA-H60 cells was compared with that of B6 NK cells cocultured with RMA (□) or RMA-H60 (▪). The graph depicts the mean (±SD) of triplicate determinations at various E/T ratios. The results shown are representative of 2 independent experiments performed.