Notch-1 Signaling Regulates Microglia Activation via NF-κB Pathway after Hypoxic Exposure In Vivo and In Vitro (original) (raw)
Figure 6
Notch blockade altered protein expression of inflammatory cytokines, iNOS and nitric oxide (NO) secretion in hypoxic BV-2 cells.
(A and B) Western blotting of TNF-α, IL-1β and iNOS (A); TGF-β1, M-CSF and IL-10 (B) protein expression in BV-2cells following 8 h of hypoxic exposure and DAPT pretreatment. The upper panel shows specific bands of TNF-α (25.6 k Da), IL-1β (17 kDa), iNOS (130 kDa) and β-actin (43 kDa) (A); TGF-β1 (25 kDa), M-CSF (18.5 kDa), IL-10 (17 kDa) and β-actin (43 kDa) (B). The lower panel in A and B are bar graphs showing significant changes in the optical density in protein expression of different groups. Note the decrease in TNF-α, IL-1β and iNOS expression (A) as well as TGF-β1 and M-CSF expression (B) in hypoxia+DAPT group compared with hypoxic BV-2 cells. A noteworthy feature was the increase in IL-10 protein expression after DAPT pretreatment in hypoxic BV-2 cells (B). (C) NO production in supernatant of different groups of cells. Note the NO production, which is increased after hypoxic exposure for 8 h is decreased nearly to basal level after hypoxic exposure in the DAPT treated BV-2 cells. Significant difference between control vs hypoxia groups is shown as *p<0.05 and **p<0.01; significant difference between control vs hypoxia and hypoxia vs hypoxia+DAPT groups is shown as #p<0.05 and ##p<0.01. The values represent the mean ±SD in triplicate.