Fig. 4.

rPKM2 increased migration factors and migration of neural progenitor cells. In vitro assays were performed to examine the neural progenitor cells (NPCs) dissected from the SVZ region. A The protein level of integrin β1 and FAK activation was examined in NPC cultures using Western blotting after 24 and 48 h exposure to rPKM2 (0.4 and 4 nM). B Quantification of Western blot data showed that NPCs exposed to 24 h of rPKM2 (4 nM) expressed a higher level of integrin β1. C The level of FAK phosphorylation (p-FAK) increased in NPCs exposed to 48 h of rPKM2 (0.4 and 4 nM). 3 replicates in each group and experiments were repeated 3 times. Asterisk, p < 0.05 versus control group. D and E Immunochemistry staining showed that these NPCs expressed immature neuronal markers Nestin (green) and DCX (red). F Transwell migration assay was performed to detect the effect of rPKM2 on the migration of NPCs. NPCs were treated with rPKM2 at different concentrations for 48 h prior to the migration measurement. 24 h after plating in transwell inserts, NPCs of the bottom membrane of inserts were fixed and stained with nuclei marker Hoechst 33342 and cytoskeleton marker Acti-stain 555 phalloidin. rPKM2 (0.4–4 nM) was added in the inserts (upper chamber) and the chemoattractant factor SDF-1 was added to the bottom inset to attract cell migration. There were more NPCs migrated to the bottom membrane of the inserts with 0.4 and 4 nM rPKM2. n = 3 independent assays; asterisk, p < 0.05 versus control group