a kit (rpa ii  (Thermo Fisher)

Journal: The Journal of Pharmacology and Experimental Therapeutics

Article Title: Prevention of HIV Protease Inhibitor-Induced Dysregulation of Hepatic Lipid Metabolism by Raltegravir via Endoplasmic Reticulum Stress Signaling Pathways

doi: 10.1124/jpet.110.168484

Figure Lengend Snippet: Effect of raltegravir and ritonavir on CYP7A1 and HMG-CoA-reductase mRNA expression and bile acid formation in rat primary hepatocytes. A, rat primary hepatocytes were treated with raltegravir (25 μM) in the absence or presence of ritonavir (25 μM) for 24 h. Total RNA was isolated. The mRNA levels of cholesterol 7α-hydroxylase (CYP7A1) and HMG-CoA-reductase were determined by RPA analysis. Rat cyclophilin mRNA was used as loading control. B, rat primary hepatocytes were treated with different amounts of raltegravir (0–25 μM) in the absence or presence of ritonavir (25 μM) for 48 h. Bile acid biosynthesis was measured as conversion of [14C]cholesterol to 14C-labeled bile acids. Labeled bile acids were extracted from media by using the Folch method. Data are expressed as a percentage of vehicle control group. The values are means ± S.E. of three independent experiments. *, p < 0.05, statistical significance relative to vehicle control.

Article Snippet: RNAqueous total RNA isolation kit and MAXIscript T7 and ribonuclease protection assay (RPA) II kits were from Ambion (Austin, TX).

Techniques: Expressing, Isolation, Labeling