Identification and Characterization of Cyclic AMP Response Element-Binding Protein H Response Element in the Human Apolipoprotein A5 Gene Promoter
Figure 1
Knockdown of CREBH expression attenuates APOA5 mRNA expression in HepG2 Cells. (a) HepG2 cells were infected with adenovirus USi (Ad-USi, 50 multiplicity of infection, MOI) or CREBH shRNA (Ad-CREBHi, 10 and 50 MOI) for 48 h. Total RNA was isolated from cells for semiquantitative PCR analysis of human CREBH and APOA5 mRNA levels and were normalized to β-actin expression. All experiments were performed in triplicate, and data represent one of three separate experiments. (b) Total RNA was isolated for Q-PCR analysis of CREBH and APOA5 mRNA levels. Data show the relative mRNA expression of CREBH shRNA (CREBHi, 10 and 50 MOI) treated samples compared to the control USi (Ad-USi, 50 MOI) treated samples. The *() and **() indicate statistically significant differences between CREBHi and USi. (c) The effects of siRNAs on CREBH and APOA5 expressions were measured by western blot analysis. β-actin expression was used as a loading control. Arrow-heads indicate the CREBH full length (upper) and CREBH active form (lower). (d) Gene expression analysis involved in lipogenesis, triglyceride synthesis, and fatty acid oxidation. Total RNAs from HepG2 cells infected with Ad-USi (50 MOI, open bar) or Ad-CREBHi (50 MOI, filled bar) were subjected to Q-PCR analysis of gene expression. mRNA levels are shown by comparing to the HepG2 cells infected with Ad-USi. *() and **() indicate statistically significant difference between CREBHi and USi.
