Skip to main content
. 2013 Jan 2;33(1):244–258. doi: 10.1523/JNEUROSCI.1665-12.2013

Figure 2.

Figure 2.

FoxP2 knockdown decreases neurogenesis in the embryonic cortex in vivo. A, Fluorescence micrographs of coronal sections through E16/17 cortices that were coelectroporated at E13/14 with nuclear EGFP and control shRNA (Control) or FoxP2 shRNA #2 (shFoxP2 #2) and immunostained for EGFP (green) and Tbr1 (red; blue is Hoechst 33258 nuclear counterstain). Tbr1 staining was used to mark the border between the intermediate zone (IZ) and the CP. In each set the left side shows the merged images of Tbr1 and Hoechst 33258, and the right side shows the EGFP immunostaining. The top white line demarcates the border between the CP and IZ, and the bottom white line the border between IZ and the VZ/SVZ. Scale bar, 100 μm. B, Quantification of sections similar to those in A for the proportion of EGFP-positive cells in the VZ/SVZ (left pair), IZ (middle pair), and CP (right pair). ***p < 0.001; n = 5 each for control and FoxP2 shRNA#1; n = 4 each for control and FoxP2 shRNA#2. C, Confocal micrographs of coronal E16/17 cortical sections immunostained for Satb2 (red) and Pax6 (middle, green) or Tbr2 (right, green). Scale bar, 50 μm. D, Higher magnification confocal images of sections similar to those shown in C. Arrows denote cells that are positive for Satb2 (red) and arrowheads denote cells that are positive for Pax6 (green, top) or Tbr2 (green, bottom; right side shows the merges). Note the absence of colocalization between Satb2-positive cells and Pax6-positive or Tbr2-positive cells. Scale bar, 20 μm. E, Confocal micrographs of coronal cortical sections from E16/17 brains 3 d after electroporation with EGFP and control shRNA (Control) or FoxP2 shRNA #2 (shFoxP2), immunostained for EGFP (green) and Satb2 (red). The white lines demarcate the different regions of the embryonic cortex. Scale bar, 50 μm. F, High-magnification confocal images of sections similar to those in E. The bottom shows Satb2 (red) alone, and the top shows Satb2 plus EGFP (green). Arrows denote EGFP-positive Satb2-positive cells. Scale bar, 20 μm. G, Quantification of sections similar to those in E and F for the percentage of EGFP-positive cells that are also positive for Satb2 or HuD, as indicated, 3 d after electroporation with control shRNA or with FoxP2 shRNA#1 or FoxP2 shRNA#2. FoxP2 shRNA#2 was somewhat less efficacious than FoxP2 shRNA#1 in this regard. *p < 0.05; **p < 0.01; ***p < 0.001; n = 3–5 embryos for each of the shRNAs. Error bars indicate SEM.