FIGURE 1.

HDAC2 activates cap-dependent translation of luciferase gene. A, overexpression of PIAS and HDAC2 on Renilla luciferase mRNA (LUC mRNA) and protein expression. The HCT-116 cells were transfected with individual PIAS or HDAC2 along with luciferase cDNA reporter gene. The cells were lysed, and the luciferase activity was measured using a luciferase assay kit (Promega). The absolute relative light units values are in the 10⁷-10⁸ range. Reverse transcription-PCR was performed using cytosolic mRNAs. To minimize the interference resulting from unequal mRNA input, the primers for β-actin and the primers for luciferase were added to the same reaction tube. VC, vector control. Error bars indicate S.E. B, HDAC2 activates cap-dependent protein translation of luciferase protein in HCT-116 cells. Expressions of HDAC isoenzymes were verified by immunoblotting. C, knockdown of HDAC2 but not of HDAC3 inhibits luciferase expression in HCT-116 cells. Protein expression of luciferase was evaluated by immunoblotting with antibody against luciferase. shRNA knockdown of HDAC2 and HDAC3 were verified by immunoblotting. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein was used a loading control.