A Reliable Primer for Detection of iNOS Expression

Part of our study was to investigate whether ATX and LPA5 antagonism would affect LPS-induced neuroinflammation in vivo. Mice (n = 6–8 per group) were injected intraperitoneally (i.p.) with DMSO, LPS (5 mg/kg) plus DMSO, with or without PF8380 (‘PF’; 30 mg/kg) or AS2717638 (‘AS’; 10 mg/kg). After 24 h, the animals were sacrificed and the brains were perfused. The right hemisphere from each mouse was processed for RNA isolation and gene expression of iNOS was evaluated by qPCR. HPRT was used as a housekeeping gene. Expression was calculated using the 2-ddCt method.

For SYBRO Green-based expression analysis of individual genes, Qiagen primers are one of the best. They are a bit pricey compared to other primers though the majority of them, including the one described here, work seamlessly, giving reproducible results.