Abnormal detachment and subsequent heterotopia formation is partially rescued with EpoD treatment. (A) Representative images of Pax6 labeling for WT and Eml1 cKO in saline or EpoD conditions at E13.5. (B and C) Quantification of the cortical wall and ventricular zone (VZ) thickness and total count for Pax6+ cells and distribution outside of VZ in WT and Eml1 cKO from saline and EpoD conditions represented as mean ± SEM (n = 6 individuals from 2 litters at least, indicated by dots of different colors). (D) Representative images of the heterotopia volume in 3D visualized by SatB2 immunofluorescence. The homotopic cortex is depicted in transparency (purple) and the heterotopia is shown with a solid rendering (red). Eml1 cKO embryos received saline or EpoD at E11.5 and E12.5 and were analyzed at E18.5. Three different angles are shown. (E) Quantification of the ratio between heterotopia volume and that of the homotopic cortex in Eml1 cKO with saline or EpoD, represented as mean ± SD (n = 7 embryos from 2 litters). Two independent litters are color-coded. (F) Quantification of Satb2 mean fluorescence intensity in the homotopic cortex in Eml1 cKO with Saline or EpoD, expressed as mean ± SD. Tests and significance: Two-way ANOVA, Sidak multiple comparison (Pax6 analyses, CW and VZ thickness. data passed normality test), Mann Whitney test (heterotopia volume and Satb2 analyses). n = 7 samples from 2 litters. P value <0.05*, <0.01 **, <0.001 ***, <0.0001 ****. Scale bars: 50 µm in A (equivalent for WT and cKO, all conditions) and 500 µm in D.