Figure 8.

Effect of rilpivirine, etravirine, efavirenz, nevirapine, and delavirdine on transactivation of the LBD of hCAR-SV23 and hCAR-SV24, and recruitment of SRC1 to the ligand-binding domain of hCAR-SV23 and hCAR-SV24. LBD-transactivation assays were performed in HepG2 cells were transfected with pCMV6-XL4-hRXRα, pFR-luc reporter plasmid, pGL4.74 [hRluc/TK] internal control plasmid, and either a receptor expression plasmid [pM-hCAR-SV23-LBD (A) or pM-hCAR-SV24-LBD (B) ] or an empty vector (pM) for 24 h. In the mammalian two-hybrid assays, HepG2 cells were co-transfected with a pM-hSRC1-RID coactivator expression plasmid, pCMV6-XL4-hRXRα, pFR-luc reporter plasmid, pGL4.74 [hRluc/TK] internal control plasmid, and either a receptor expression plasmid [pVP16-hCAR-SV23-LBD (C) or pVP16-hCAR-SV24-LBD (D) ] or an empty vector (pVP16) for 24 h. Transfected cells were treated with DMSO (0.1% v v⁻¹; vehicle control), rilpivirine (RPV; 5 μM), etravirine (ETV; 5 μM), efavirenz (EFV; 5 μM), nevirapine (NVP; 5 μM), or delavirdine (DLV; 5 μM), TCPOBOP (0.25 μM; negative control), DEHP (10 μM; positive control for hCAR-SV23), or CITCO (10 μM; positive control for hCAR-SV24) for 24 h. Firefly luciferase and Renilla reniformis luciferase activities were measured and normalized as described under Methods. Data are expressed as mean ± SEM for three independent experiments performed in triplicate. *Significantly different from the same treatment group transfected with corresponding empty vector and from the vehicle-treated control cells transfected with respective receptor expression plasmid (P < 0.05).