FIGURE 4.

Effect of CHOP overexpression on endogenous gene expression. HEK293T cells were transfected with GFP-pcDNA3.1 or CHOP-pcDNA3.1 plasmids as described under “Materials and Methods.” A shows phase contrast fluorescence images of the GFP-transfected cells (2 μg/60-mm dish). At 24 h post-transfection, cells were incubated for 8 h in MEM or in MEM containing 2 mm HisOH, and then RNA and protein extracts were isolated. Protein extracts from GFP- or CHOP-transfected cells were subjected to immunoblot analysis to detect the presence of CHOP protein (B). The RNA was subjected to qRT-PCR analysis for the mRNA content of the indicated genes, as well as GAPDH mRNA as an internal control. The results are shown as the means ± S.E. for three independent experiments (C). An asterisk indicates that the induction of expression was significantly changed by CHOP expression (p < 0.05).