Figure 4

MOLT-13 and MOLT-13 boost DNA and cDNA sequencing analysis. (A) MOLT-13 TP53-sequencing. 1. DNA analysis shows a heterozygous mutation in codon 273 (TP53, exon 8, CGT > CAT, Arg > His) and a heterozygous polymorphism in codon 72 (TP53, exon 4, CGC > CCC, Arg > Pro), identical ratio of wild type to mutated template was detected. 2. cDNA sequencing confirms DNA results. (B) MOLT-13 boost TP53-sequencing. 1. DNA analysis shows a heterozygous mutation in codon 273 and, surprisingly, a deletion of one nucleotide - a nonsense mutation - in exon 4 (both wild type and mutated template are visible). 2. cDNA analysis showed only mutated nucleotide in codon 273 (representing hemizygous mutation) and only one allele expression in exon 4. These results suggest a missense mutation of one allele and a nonsense mutation of the other causing the nonsense mRNA decay. (C) FBXW7 and N-RAS DNA sequencing. In order to confirm MOLT-13 cell line identity, mutations of FBXW7 and N-RAS gene were verified. 1. FBXW7 analysis shows a heterozygous mutation in codon 465 (FBXW7, CGT > CAT, Arg > His). 2. N-RAS sequencing shows a heterozygous mutation in codon 12 (N-RAS, GGT > GAT, Gly > Asp). Mutations are marked with arrows. N - normal, control template.