| Popis: |
Supplementary Figure S1: Breeding/Genotyping of mouse lines used in this study. (A, B) Breeding crossings used. K14-Cre'g/O I RxraL2/L2 is also known as RXRaep-/-. (C) Semi-quantitative PCR was performed on biopsies containing both epidermal and non-epidermal tissue; to confirm genotypes with regard to presence/absence of K14-Cre recombinase (top), epidermis-specific ere-mediated excision of Rxra exon 4 (middle), and the mutant Cdk4R24c allele (bottom). Presence of K14-Cre recombinase is indicated by a single 349 bp band; animals negative for K14-Cre show no amplification. In K14-Cre positive animals, ere-mediated epidermis-specific excision of Rxra is indicated by a 110 bp (L-) band, while a 203 bp (L2) band is also amplified due to presence of non-epidermal tissue in the biopsy in which excision did not occur. Animals negative for K14-Cre show only the 203 bp (L2) band for Rxra, as no excision occurred in any tissue. Animals homozygous for the mutant Cdk4R24c allele show a single band at 530 bp, while animals homozygous for the WT Cdk4 allele show a single band at 440 bp. Primers used are detailed in the Supplementary Table S2. (D) Tyr-NRAS061K genotype is determined phenotypically 10-12 days postnatal by presence/absence of dark pigmentation of the foot pads. |
