Figure 5
K-181 exerts neuroprotective effects in response to hypoxia/reperfusion in neurons. (A,B) Representative western blot images (A) and quantification of pMdmx/Mdmx relative expression (B) in neurons which were pre-treated with DMSO and 10 μM of K-181 after 3 h OGD following 9 h reperfusion. N = 3. (C,D) Representative immunostaining images (C) and quantification (D) of Mdmx in neurons, before and after hypoxia/recovery. N = 6. Scale Bar = 100 μm. (E) Cell death rate of neurons which were pre-treated with DMSO, K-181 and SJ-172550 at 24 h after 3 h OGD. N = 4–6. (F,G) Representative immunostaining images (F) and quantification (G) of phosphorylated cyclic AMP-responsive element-binding protein (p-CREB) in neurons, before and after hypoxia/recovery. N = 6. Scale Bar = 100 μm. (H) Western blot of p53 in neurons transduced with shRNA-control and shRNA-p53. (I) qPCR of p53 mRNA in neurons transduced with shRNA-control and shRNA-p53. N = 3. (J) Cell death rate of neurons which were transfected with shRNA-control and shRNA-p53 at 12 h after 3 h OGD. N = 4–6.
