Extended Data Fig. 1: Validation of Trp53 mCherry and Trp53 R172H-Akaluc alleles.

(a) Genotyping PCR for Trp53 wildtype, PAL/ + , PAL/PAL mice using F2/R2/R3 primers in Fig. 1a. 240 bp, wildtype band. 416 bp, mutant band. Results are representative of at least 80 independent genotyping experiments. (b) Southern blot of EcoNI- or ScaI-digested tail DNA from Trp53 PAL/+ mice using 5’ probe or RR probes in Fig. 1a. Trp53 PAL shows specific 11.1 kb or 3.5 kb bands, respectively. Results are representative of 8 independent experiments. (c) Genotyping PCR for Trp53 mCherry/+ mice using CF1/R1 or CF2/R2 primers in Fig. 1a. Trp53 wildtype mouse was used as a control, showing no band. Results are representative of at least 80 independent genotyping experiments. (d) Southern blot of EcoNI-digested tail DNA from Trp53 mCherry/+ mice using 5’ probe or RR probes in Fig. 1a. Trp53 mCherry shows specific 5.4 kb or 4.5 kb bands, respectively. Results are representative of 5 independent experiments. (e) Targeted Sanger sequencing for PCR products from Trp53 PAL/+ tail DNA using F1 or F2/R3 primers in Fig. 1a, highlighting the G to A mutation at the R172H site, and the junction sequences between exon 11, linker, Akaluc, and 3’ UTR.