Figure 5. Nrf2 activation is required for MIND4-17-mediated cytoprotection.

OB-6 cells, stably expressing lentiviral scramble non-sense control shRNA (“sh-scr”) or Nrf2 shRNA (“sh-Nrf2, sequence -1/−2”) (A-E), as well as lentiviral Keap1 shRNA (“sh-Keap1”) (F-J), were treated with MIND4-17 (3 μM) for 3 hours, expressions of listed proteins were presented (A and F); mRNA expressions of listed genes were also shown (B, C, G and H); Cells were further treated with/out hydrogen peroxide (“H₂O₂”, 200 μM) for indicated time period, cell viability (CCK-8 assay, D and I) and apoptosis (Histone DNA ELISA assay, E and J) were also tested. For H₂O₂ experiments, MIND4-17 (3 μM) were pretreated before H₂O₂ for 1 hour. For all the assays, the exactsame number of viable cells with the applied shRNA was initially plated into each well. Data were presented as mean (n=5) ± standard deviation (SD). ^#p<0.05. Experiments in this figure were repeated for three times, and similar results were obtained.