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. 2013 Aug;94(2):301–313. doi: 10.1189/jlb.0213093

Figure 2. KIR2DS4 surface expression is decreased after knockdown of DAP12 in primary cells.

Figure 2.

Primary cells were treated with nontargeting (nt) or DAP12 targeting (t) siRNA. (A) The relative quantity of DAP12 mRNA transcripts was determined by rqRT-PCR using β-actin as an internal loading control from RNA isolated from PBMCs or KIR2DS4 positively sorted cells from these PBMCs. The relative quantity of DAP12 mRNA was normalized to levels expressed following treatment with the nontargeting siRNA. The data represent the mean (±sem) from five independent experiments performed in duplicate from two individuals. (B) Representative Western blot of DAP12 and densitometry analysis of PBMC whole cell lysates following treatment with nontargeting or DAP12-targeting siRNA. The density values obtained for β-actin were used to normalize the DAP12 density values. The data represent the mean (±sem) from five independent experiments performed in duplicate from two individuals. (C) KIR2DS4 surface expression from two donors was normalized to expression levels obtained following nontargeting siRNA treatment for whole PBMCs or KIR2DS4 positively sorted cells. The results were recorded from five independent experiments performed in duplicate and are represented by the normalized mean ± sem. Statistical analyses for all of the primary cell experiments were performed with a Mann-Whitney U-test (*P<0.05; **P<0.01).