Research Paper Volume 11, Issue 24 pp 12032—12042

Compound 13 activates AMPK-Nrf2 signaling to protect neuronal cells from oxygen glucose deprivation-reoxygenation

Figure 5. C13 activated AMPK downstream Nrf2 signaling in neuronal cells. SH-SY5Y human neuronal cells (AC) or the primary murine hippocampal neurons (D and E) were treated with applied concentration of C13 for indicated time, expression of listed mRNAs (A and E) and proteins (in both total lysates and nuclear fraction lysates, BD) were shown. 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 indicated time, expression of listed mRNAs were shown (F and G). The stable SH-SY5Y cells with Nrf2 shRNA (“sh-Nrf2” cells) and CRISPR/Cas-9 Nrf2-KO construct (“ko-Nrf2” cells), as well as the parental control cells (“Pare”), were treated with C13 (10 μM) for indicated time, listed mRNAs were shown (H and I); Cells were also pretreated with C13 (10 μM) for 2h, followed by OGD-R stimulation for 36/48h, cell viability (CCK-8 assay, J) and apoptosis (Histone DNA ELISA assay, K) were tested. Expression of listed proteins were quantified and normalized to the loading controls (BD). Bars stands for mean ± standard deviation (SD, n=5). * p<0.05 vs. “Vehicle” control cells (A and E). #p<0.05 vs. “Pare” cells (F-K). Each experiment was repeated three times with similar results obtained.