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. Author manuscript; available in PMC: 2025 Aug 25.
Published in final edited form as: J Cell Biochem. 2023 Aug 29;124(10):1530–1545. doi: 10.1002/jcb.30462

FIGURE 2.

FIGURE 2

Electrical stimulation increased neurite outgrowth and DNA demethylation in PC-12 cells. (A–C) ES increased neurite outgrowth in PC12 cells compared to control unstimulated cells. PC12 cells incubated with a low level of NGF (5 ng/mL) (panels 1 and 2) and received 20 min ES at 24 h post NGF induction. (D–F) ES increased neurite outgrowth in PC12 cells compared to control unstimulated cells. PC12 cells incubated with a low level of NGF (5 ng/mL) (panels 1 and 2) and received 20 min ES at 24 h post NGF induction. However, pre-incubating PC12 cells with TETi76 diethyl ester (a pan TET inhibitor) at 12.5 μM (for 16 h) before ES (panel 3), blocked the ES induced neurite outgrowth. Result of qPCR assessing mRNA levels of GAP43 (G) and TET1 (H) expression in Non-ES- and ES-treated cells. (I) Immunocytochemistry based analysis of 5hmc (green) expression, an intermediate product of DNA demethylation process, reveals increased expression of the substrate following ES. The expression was localized in the nucleus (blue). Values are means ± SEM. **p ≤ 0.01; *p ≤ 0.05 by the Student t test. mRNA, messenger RNA; NGF, nerve growth factor.