Fig. 8

Clemastine inhibits expression of IL-1β and NLRP3 and exerts no effect on the expression of HH1R of microglial cells in vitro. Double immunofluorescence staining showing the distribution of Iba-1-labeled (a, d and g, green) and HH1R (b, e and h, red) immunoreactive AMCs in vitro at 1 h after OGD and OGD + clemastine and their corresponding controls. The colocalized expression of Iba-1 and HH1R in AMCs can be seen in c, f, and i. Panels j and n show bar graphs depicting significant increases in the immunofluorescence intensity and optical density of HH1R following the OGD when compared with their corresponding controls, clemastine and MCC950 showed no effect on the expression of HH1R. Panel k showing IL-1β (17 kDa), NLRP3 (62 kDa), HH1R (70 kDa), and β-actin (42 kDa) immunoreactive bands. Panels k and l are bar graphs which indicate that expression of IL-1β and NLRP3 were improved in the AMCs after OGD administration for 1h. The clemastine and MCC950 may attenuate the increment. N=3. *P < 0.05. Scale bars: a–i 50μm