Figure 6. Expression level of C/EBPϵ mRNA in various cell lines, and TNFα-induced up-regulation of PHGPx mRNA expression in HEK-293 cells overexpressing C/EBPϵ.

(A) Total cellular RNA was isolated from the cells, and the mRNA levels of C/EBPϵ, CREB and NF-Y were detected by RT-PCR. After PCR, 10 μl of the PCR products were subjected to electrophoresis and the DNA was visualized by ethidium bromide staining. Reproducibility of these results was confirmed in three experiments. (B) HEK-293 cells transiently overexpressing a transcription factor (NF-Y, CREB, NF-κB, C/EBPϵ, C/EBPδ or C/EBPγ) were incubated for 24 h with (black bars) or without (grey bars) 50 ng/ml TNFα. Total cellular RNA was isolated from the cells, and mRNA levels of PHGPx and 18S rRNA as an internal control, were detected by quantitative PCR. Results are means±S.D. of the percentage of the vector control incubated without TNFα. Reproducibility of these results was confirmed in two experiments.