Fig. 5.

Inhibition of SUMOylation enhances RXRα target gene expression. Each graph shows a representative experiment, performed in triplicate, of a minimum of three experiments. (A) HuH-7 cells were pretreated with vehicle, 10 μM GA, or 50 μM AA for 4 hours prior to CDCA (if applicable) for an additional 20 hours. cDNA was made from RNA extractions, and real-time polymerase chain reaction was performed with primers for each listed gene. All expression, except glyceraldehyde 3-phosphate dehydrogenase (GAPDH), is relative to 18s and normalized to vehicle-treated cells. (B) HuH-7 cells were transfected with empty vector, RXRα, or RXRαK108R (SUMOylation deficient) plasmids and treated with vehicle or 9cRA 24 hours prior to harvest and measuring of luminescence. Data are normalized to empty vector, vehicle-treated cells. (C) HuH-7 cells were pretreated with vehicle and 10 μM GA for 4 hours prior to 9cRA (if applicable) for an additional 20 hours. cDNA was made from RNA extractions, and real-time polymerase chain reaction was performed with primers for each listed gene. All expression, except GAPDH, is relative to 18s and normalized to vehicle-treated cells. Data are presented as mean + S.E. and *P < 0.05; **P < 0.01; and ***P < 0.001.