Fig. 4

Loss of Gli2 mutant GCPs at P0 is compensated by wild type (WT) cells at P8. a-b In situ RNA hybridization analysis of Gli1 on P8 midsagittal cerebellar sections of Gli2^lox/lox (control, a) and Atoh1-Cre/+; Gli2^lox/lox (Atoh1-Gli2 CKO, b) mice. Red arrowhead indicates the strong Gli1 expression in the mutant EGL and red asterisks indicate normal Gli1 expression in Bergmann glia in the Purkinje Cell Layer (PCL). c-d FIHC detection of the indicated proteins and dapi on P8 mid-sagittal cerebellar sections of Gli2^lox/lox (control, c) and Atoh1-Cre/+; Gli2^lox/lox (Atoh1-Gli2 CKO, b) mice. High power images are shown of the areas indicated by white rectangles in (c and d) with the thickness of the outer EGL (o) indicated by yellow brackets. The white arrow in (b) indicates the proliferating EGL. e and f Graphs of the thickness proportion of the EGL at P8 (n = 4) (e) and the proportion of oEGL area at P8 (% [oEGL] area of total [EGL] area) (n = 4) (f) in Gli2^lox/lox (CTL, black) and Atoh1-Cre/+; Gli2^lox/lox (Atoh1-Gli2 CKO, red) mice. All of the analyses were performed on 3 midline sections per brain. All graphical data are presented as means ± SEM and significance determined using two-tailed T-test. g-h In situ hybridization of Cre RNA on P8 midsagittal cerebellar sections of Gli2^lox/lox (control, g) and Atoh1-Cre/+; Gli2^lox/lox (Atoh1-Gli2 CKO, h) mice. Black arrows indicate the loss of Cre expression in the partially rescued EGL. i-k FIHC detection of the indicated proteins and dapi on P8 mid-sagittal cerebellar sections of Gli2^lox/lox (control, j) and Atoh1-Cre/+; Gli2^lox/lox (Atoh1-Gli2 CKO, i and k) mice. EGL is indicated by the yellow dotted lines and yellow asterisk indicates low level of SOX2 expression in the mutant EGL . Scale bars represent 100 μm