Implication of PGC-1α in sPLA2-IIA promoter activity.

A/ Implication of PGC-1α in sPLA2-IIA promoter activity. 158N cells were transiently transfected with either 0.2 µg of sPLA2-IIA(1 kb), 0.1 µg of pRSV-βGal plasmids with PGC-1α expression vector, with mock vector or with a siRNA specifically directed against PGC-1α, as indicated. Eighteen hours after transfection, cells were incubated with 25-OH (10−5 M) for 24 h, and then luciferase and β-galactosidase activities were analyzed. Results are expressed as percentage of the basal activity and they represent the mean +/− SEM of at least four independent experiments performed in duplicate. *p<0.05 when compared between control cells and cells transfected with PGC-1α expression vector or siRNA against PGC-1α using Bonferroni's test after ANOVA. B/ The efficacy of the siRNA was analyzed by RT-PCR. 158N cells were transiently transfected with a siRNA specifically directed against PGC-1α. 24 hours after transfection, total RNA was prepared. RT-PCR experiments were performed using primers recognizing specifically PGC-1α. PCR products were analyzed on agarose gel (2%) and visualized under UV. 18S RNA was detected by specific primers and used to normalize PGC-1α expression levels.