Figure 5

Effects of the combination of ACR and LY294002 on the phosphorylation of RXRα, ERK, and Akt proteins and the transcriptional activity of the RXRE promoter in HLF cells. (A) The cells were treated with vehicle, 1 μM ACR alone, 5 μM LY294002 alone, or a combination of 1 μM ACR and 5 μM LY294002 for 12 hours. The extracted proteins were examined by western blot analysis using the respective antibodies. Repeat western blots gave similar results. (B) A transient transfection reporter assay was performed with the RXRE luciferase reporter in the presence of vehicle, 1 μM ACR alone, 5 μM LY294002 alone, or a combination of 1 μM ACR and 5 μM LY294002. Relative luciferase activity was determined after 24 hours. Columns and lines indicate the means and SD of triplicate assays. # P < 0.01. * P < 0.05.