Figure 1.

In vitro validation of anti-rCCR2 DsiRNAs. Ten 27-mer DsiRNA duplexes against rCCR2 were transfected in HEK293 cells stably expressing the rCCR2 receptor. Cells were treated with increasing doses of DsiRNAs complexed with the delivery reagent RNAiMAX, 24 h before RNA extraction. rCCR2 mRNA quantification was then assessed by quantitative real-time PCR. As a control, cells were also treated with RNAiMAX alone (vehicle group). Knockdown levels of CCR2 were normalized to levels of internal control RLPO and HPRT and normalized to the negative mismatch control = 100%. Duplexes 5 and 6 were considered as the most potent and were used for further optimization. Bars represent mean ± SEM, n = 3.