Fig. 1. (A) Generation of inducible Gli1CreERT2/+:: R26RYFP/+ mice. (A) Breeding scheme for generation of Gli1CreERT2/+:: R26RYFP/+ mice to confirm the Cre activity according to tamoxifen induction. (B) Schematic experimental design for the tamoxifen injection time points used in Gli1CreERT2/+:: R26RYFP/+ mice. Pups were orally injected TAM on P4~7. Pups were sacrificed to collect the brain and tail samples at different indicated time points. (C) Representative genotyping results of offspring from R26RYFP/YFP reporter and Gli1CreERT2/+ crosses. Genomic DNA (gDNA) from brain regions (cerebellum and cortex) and tail were examined by PCR for recombination of floxed alleles in Gli1CreERT2/+:: R26RYFP/+ mice. Recombinant ΔR26R-YFP, which indicates the transgene of cre-mediated recombination driven by Gli1 activation was observed in cerebellum but not in cerebral cortex and tail of Gli1CreERT2/+:: R26RYFP/+ mice. Mice not expressing Cre from parallel breeding were taken as control (lane 1). (D~D”) Sagittal whole-brain images from tamoxifen injected Gli1CreERT2/+:: R26RYFP/+ mouse at P25, indicating that Cre-recombinase is selectively expressed in the cerebellum. Green fluorescence, YFP signal indicates Cre-mediated recombination. Counterstaining was performed using Bisbenzimide (Hoechst 33342) dye. (D’) YFP signal was dominantly detected in cerebellar cortex of Gli1CreERT2/+:: R26RYFP/+ mice. Boxed region denotes enlarged area in D”. D” showed each layer of cerebellum expressing YFP signal. P, postnatal day; TAM, tamoxifen; IHC, Immunohistochemistry; CB, cerebellum; CTX, cerebral cortex; OB, Olfactory bulb; HP, Hippocampus; TH, thalamus; HY, Hypothalamus; MB, Midbrain; P, Pons; M, Medulla; ML, Molecular layer; PCL, Purkinje cell layer; GCL, Granule cell layer. Scale bars=1 mm in D and D’, 0.2 mm in D”.
© Exp Neurobiol
