The length of the PCR products of VIP, VPAC1, NKG2D, DAP10, NF-[kappa]B, and beta-actin was 361 bp, 432 bp, 365 bp, 282 bp, 326 bp, and 500 bp, respectively.
After slides dried, the target proteins of NK cells were detected by immunocytochemistry test according to the procedure of the PV-9000 kit (for detecting VIP, DAP10, VpAc1, and NF-[kappa]B) or PV-6003 kit (for detecting NKG2D) (Zhong Shan Company, Beijing, China).
The Expressions of NKG2D, DAP10 and NF-[kappa]B p65 mRNA and Protein.
VIP antagonist could partially or completely abolished the effect of VIP on the expressions of NKG2D, DAP10 and NF-[kappa]B in NK cells (P < 0.05, Table 6, Figures 4, 5, 6 and 7).
NKG2D signal transduction should through two adaptor proteins, DAP10 and DAP12 , which were associated with the receptor as homodimers.
In our study, we found that VIP downregulated the NKG2D, DAP10, and NF-[kappa]B expression in NK cells, which could be reversed completely or mostly by VIP antagonist.