The pMN expansion is restored over time when Arl13b is deleted at E9.25 and E9.5. (A-D) Olig2 (red) and HB9 (green) cells are expressed in the pMN domain of control at E10.5 (A), but are expanded in Arl13bhnn (B), Arl13bΔE9.25 (C) and Arl13bΔE9.5 (D). (E-H) Shh (green) is expressed in the notochord and the floor plate of E10.5 control (E) and Arl13bΔE9.5 (H), but only in the notochord of Arl13bhnn (F) and Arl13bΔE9.25 (G). (I-L) At E12.5, there is normal Olig2 (red) and HB9 (green) expression in control (I) and Arl13bΔE9.5 (L), whereas there is an expansion of cells in Arl13bhnn caudal neural tube (J). Olig2 and HB9 cells are mainly in their correct domains in Arl13bΔE9.25, except that there are some dorsally expressed Olig2 and HB9 cells (K and white arrows). (M-P) In control (M), Arl13bΔE9.25 (O) and Arl13bΔE9.5 (P) caudal neural tube, Shh is expressed in the notochord and floor plate at E12.5, whereas it is expressed in only the notochord of Arl13bhnn neural tube (N). (Q) Quantitative results showing that the percentage of normalized Olig2 (red) and HB9 (green) cells in E12.5 Arl13bΔE9.25 and Arl13bΔE9.5 caudal neural tube are similar to control, whereas there are excessive Olig2 and HB9 cells in Arl13bhnn. Data are mean±s.d. *P<0.05.