Figure 5.

Cell number in the RPE layer is increased in the Nf2^CKO dorsal optic cup at E11.0. (A, B) MITF labeling reveals increased RPE layer thickness in Nf2^CKO (B; arrowheads point to ectopic RPE cells located apically). (C, D) Quantification indicates a significant increase in total cell number in the dorsal RPE in Nf2^CKO (C; see Supplementary Material, Figure S3C for subdivisions; control: 183.75 ± 15.13 SD; Nf2^CKO: 271.00 ± 10.42 SD), while cell number is unchanged in the dorsal retina (D). Data are means ± SD, Student’s T-test was applied for statistical analysis, and P-values are indicated on the horizontal lines in each graph (significance < 0.05). (E) Phalloidin labeling in control demonstrates regular organization. (F) Phalloidin-labeling Nf2^CKO reveals irregularities in localization in the dorsal RPE layer (arrowheads). (G) Normal localization of pMLC2 in control dorsal RPE. (H) In addition, mis-localization of pMLC2 is detectable in the dorsal RPE layer in Nf2^CKO optic cups (arrowheads). (I, J) OTX2 labeling shows that supernumerary cells persist in the dorsal optic cup at E13.5 (J; arrowheads). Scale bars: 50 μm in A, I.