Fig. 4

Double immunofluorescence staining of CSF1 and CSF1R with Iba-1 in rat brain at 48 h after HI. a Representative microphotographs of CSF1 and CSF1R and Iba-1 staining in the ipsilateral cortex of rat brain. CSF1 and CSF1R were colocalized with Iba-1 positive microglia in the sham, vehicle, and rh-CSF1 treatment groups, respectively. b Brain slice of rats at 48 h after HI. The upper right panel of brain slice indicates the location for staining analysis (small blue box). c, d Quantitative analysis of CSF1-positive Iba-1 and CSF1R-positive Iba-1 cells in the ipsilateral cortex at 48 h after HI. Compared with the sham group, immunofluorescence staining showed an increase in expression of CSF1 and CSF1R on microglia in the vehicle group and rh-CSF1-treated animals. Intranasal administration of rh-CSF1 further increased the expression of CSF1 and CSF1R on microglia than that of the vehicle group. Green indicated CSF1 and CSF1R positive staining, red indicated Iba-1 positive microglia staining, and blue indicated DAPI positive nuclear staining. Merge showed the colocalization of CSF1 and CSF1R with microglia. *P < 0.05 vs sham; ^#P < 0.05 vs vehicle; Scale bar =50 μm; n = 4 in each group