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. Author manuscript; available in PMC: 2013 May 21.
Published in final edited form as: Cell. 2012 Nov 21;151(5):1097–1112. doi: 10.1016/j.cell.2012.10.043

Figure 5. Znf335 deficiency leads to decreased cell size, and abnormal dendritic shape and orientation.

Figure 5

(A) Knockdown of ZNF335 leads to reduction of cells in the cortical plate and production of Cux1-negative (white circles) and FoxP1-positive (blue circles) cells showing a change in cell fate. Dashed line represents end of zone containing GFP-positive cells. Scale: 50μm.

(B) Knockdown and control cells were targeted at E14.5 and analyzed at P0 (a,b), P6 (c,d) to show abnormal radial glia (P0, arrowhead) and abnormal cell body shape and size (P6, dashed circle). There is abnormal dendritic arborization (arrowhead) and orientation in knockdown cells at P8 (e,f), P16 (g,h), P22 (i,j,k), and P30 (l,m,n). Scale: 50μm.

(C) Analysis of 5A. Knockdown showed production of more Cux1-negative cells as compared to Control, and production of more FoxP1-positive cells as compared to Control.

(D). Knockdown cells show abnormal orientation based on orientation of their basal dendritic process which is normally perpendicular to the pial surface of the brain. WT-ZNF335 rescues the orientation but MUT-ZNF335 does not. Scale: 50μm. 0.92 of total UT-Control cells have apical process orientated perpendicular to pial surface (0°), 0.03 at 22.5°, and 0.04 at 337.5°. shRNA-ZNF335 knockdown cells have only 0.25 of total cells oriented at 0°, 0.13 at 22.5°, 0.11 at 45°, 0.08 at 67.5°, 0.04 at 90°, 0.04 at 270°, 0.08 at 292.5°, 0.12 at 315°, and 0.13 at 337.5°. WT-ZNF335 rescue of shRNA-ZNF335 knockdown have 0.7 cells at 0 °, 0.03 at 22.5 °, 0.05 at 45 °, 0.3 at 67.5 °, 0.04 at 90 °, 0.02 at 270 °, 0.03 at 292.5 °, 0.07 at 315 °, and 0.05 at 337.5 °. MUT-ZNF335 rescue of shRNA-ZNF335 knockdown have 0.4 cells at 0 °, 0.1 at 22.5 °, 0.11 at 45 °, 0.06 at 67.5 °, 0.01 at 90 °, 0.03 at 270 °, 0.06 at 292.5 °, 0.08 at 315 °, and 0.14 at 337.5 °. Data presented as proportion of total cells oriented in +/−11.25 ° of radial direction n=6 electroporated brains. Only matching sections were analyzed between different conditions.

See also Fig S4.