Figure 3. C13 activated AMPK signaling in SH-SY5Y neuronal cells and primary neurons. SH-SY5Y human neuronal cells (A–B) or the primary murine hippocampal neurons (C and D) were treated with applied concentration of compound 13 (“C13”) for 1h, expression of listed proteins in total cell lysates were shown (A and C); The AMPK activities were also tested (B and D). The stable SH-SY5Y cells with AMPKα1 shRNA (“sh-AMPKα1”) and CRISPR/Cas-9 AMPKα1-KO construct (“ko-AMPKα1”), as well as the parental control cells (“Pare”), were treated with C13 (10 μM) for 1h, expression of listed proteins (E) and AMPK activities (F) were tested. The cells were pretreated with C13 (10 μM) for 2h, followed by OGD-R stimulation for applied time periods, cell viability and apoptosis were tested by CCK-8 assay (G) and Histone DNA ELISA assay (H), respectively. Bars stands for mean ± standard deviation (SD, n=5). * p<0.05 vs. “Mock” cells. #p<0.05 vs. parental control cells. Each experiment was repeated four times with similar results obtained.