Fig. 5

Effect of HIV-1_ADA gp120 on the protein and mRNA expression of PPARγ in vitro and in vivo. Primary cultures of rat astrocytes were exposed to g120 (5 nM) for 6 h, and PPARγ mRNA levels were measured using qPCR (a). Primary cultures of mixed rat astrocytes and microglia were exposed to g120 (5 nM) for 3 h, and PPARγ mRNA levels were measured using qPCR (b). Adult Wistar rats were administered, bilateral ICV, 4 μg/ventricle of gp120; hippocampus was isolated 6–72 h post ICV, and PPARγ mRNA levels were measured using qPCR (c), and protein expression of PPARγ was analyzed through immunoblotting (d). Cyclophillin was used as the housekeeping gene for qPCR. For immunoblotting, hippocampus tissue protein lysates (50 μg) were resolved on a 10% SDS-polyacrylamide gel and transferred to a PVDF membrane. HepG2 (50 μg) were used as positive control for PPARγ protein. PPARγ was detected using a rabbit polyclonal PPARγ antibody (1:1000 dilution). pt?>Actin was detected using a mouse monoclonal antibody (1:5000, dilution). Data generated from densitometric analysis is presented as a ratio of PPARγ expression normalized to actin (loading control). Results are expressed as mean ± SEM relative to DMSO (control, in vitro) or saline (control, in vivo) of at least 3 separate experiments in vitro and n = 6–11 animals/group in vivo. Asterisks represent data point significantly different from DMSO or saline (control) (****p < 0.0001, *p < 0.05)