Role of Reactive Oxygen Species in the Neural and Hormonal Regulation of the PNMT Gene in PC12 Cells

Figure 4

Effect of ROS on the neural regulation of the PNMT gene. PC12 cells were treated with 200 μM CoCl₂, 10 μM forskolin or the combination of CoCl₂ and forskolin for 24 h at 37°C. (a) Luciferase activity from cells transfected with the wild-type pGL3RP893 PNMT promoter luciferase reporter gene construct. (b) Representative image (𝑛=3) of PNMT (intron-retaining and intronless splice variants) mRNA levels from semi-quantitative RT-PCR. GAPDH mRNA was used as a housekeeping control. (c) PNMT intron-retaining mRNA and (d) intronless mRNA. Data is normalized to GAPDH control. Data presented as relative fold change ± SEM from a minimum of three independent experiments. Significance defined as *𝑃<0.05; ^**/++𝑃<0.01; ^+++/###𝑃<0.001.

(a) Transfection

(b) Representative gel image

(c) PNMT intron-retaining mRNA

(d) PNMT intronless mRNA