Subclones with distinct codon sequences in the 9-basepair region of the N-gly site region. (A) N-gly site status of subclones with a different codon sequence in N-gly site region to that of the major clone. N-gly site-positive subclones represented here do not display either the same asparagine or serine/threonine encoding nucleotide sequence as that of the major clone. The middle codon for all sequences was checked to ensure for the absence of proline, as this middle amino acid would negatively affect the functionality of the N-gly site. Percentages were calculated from the values in columns 5 and 6 from Table 3. (B) Patient 3 subclones from the 3 disease events are illustrated as an example. The pie charts provide an overview of the distribution of subclones harboring either synonymous (green) or nonsynonymous (orange) mutations compared with the sequence of the major clone. The numbers inside the pie charts are representative of the actual number of subclones. For the second relapse and transformation subclones, the majority of subclones have a different amino acid sequence encoding for the N-gly site (nonsynonymous), whereas the majority of the third relapse subclones maintain the amino acid sequence found in the major clone, which is NIS (synonymous). The tables highlight the variety of amino acid sequences that encode for N-gly sites (eg, NIT, NVT, NIS) found in these subclones and the diverse range of codon sequences that are responsible for both these synonymous and nonsynonymous mutations. Numbers in brackets represent the number of unique subclones presenting with the particular codon sequence.