Fig. 7: Integrin β1 is involved in Cxcr4-mediated blood vessel-guided migration.
From: Erratic and blood vessel-guided migration of astrocyte progenitors in the cerebral cortex
a Simultaneous introduction of Cxcr4 and Cxcr7 CRISPR vectors reduced the amount of Integrin β1 on astrocyte progenitors (3 brains/group, two-sided unpaired Student’s t test, P = 0.0383, the data in the right panel are shown as mean ± SEM). CRISPR vectors were electroporated at E15, and the amount of Integrin β1 on the GFP and ACSA-2 double positive population at P3 was examined using a flow cytometer. The left dot plot shows the fluorescence intensity of the ACSA-2 (y-axis) versus the intensity of forward scatter (FSC; x-axis). The right dot plot shows the fluorescence intensity of the GFP (transfected cells, y-axis) versus the intensity of forward scatter (FSC; x-axis). Histogram shows the number of events (cells) counted (y-axis) versus the fluorescence intensity of integrin β1 (x-axis) (red, ACSA-2+, GFP+ cells electroporated with a control vector; blue, ACSA-2+, GFP+ cells co-electroporated with Cxcr4 CR#1 and Cxcr7 CR#2). b Knockdown efficiency of two independent vectors (KD#1 and KD#2) for Itgb1 (n = 4 independent transfections to Neuro2a cells). c E15 embryos were electroporated with knockdown vectors and/or Cxcr4 expression vector and fixed at P3. Scale bar, 20 µm. d Distances from blood vessels to transfected astrocyte progenitors (GFP+/Aldh1l1+). Both KD#1 and KD#2 significantly increased the distances from blood vessels. Cxcr4 overexpression enhanced the association with blood vessels, while this effect could be canceled by adding KD#1 (two-sided Tukey-Kramer tests, 1929 cells/18 brains control, 691 cells/8 brains Itgb1 KD#1, 844 cells/9 brains Itgb1 KD#2, 325 cells/13 cells Cxcr4 OE, and 245 cells/7 brains Cxcr4 OE + Itgb1 KD#1, control vs. KD#1, P = 0.01406; control vs. KD#2, P = 0.00321; control vs. Cxcr4-OE, P = 0.02445; Cxcr4-OE vs. Cxcr4-OE + Itgb1 KD#1, P < 0.001). For detailed information of box plots, see “Statistical analysis” section in Methods. *p < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.
