Characterisation of PALB2 mutation transcripts via RT-PCR. (a) Image of transcripts from PALB2 c.3113G>A visualised on 3% agarose gel. (A) Heteroduplex of the RT-PCR products of the wild-type and the PALB2 r.2997_3113del transcripts. (B) Heteroduplex of the RT-PCR products of the wild-type and PALB2 r.3083_3113del transcripts. (C) RT-PCR product of the wild-type transcript. (D) RT-PCR product of the PALB2 r.3083_3113del transcript. (E) RT-PCR product of the PALB2 r.2997_3113del transcript. FSIs of six wild-type and their corresponding six variant nucleotides at heterozygous positions were recorded for each condition (CT and NCT). Averaged proportions of FSI of the six variant nucleotides to the FSI of the six corresponding wild-type nucleotides for each condition are shown. (b) Image of wild-type transcripts and transcripts from PALB2 c.196C>T visualised on 4% agarose gel. FSIs were recorded for the variant and corresponding wild-type nucleotide at the heterozygous position. (c) Image of transcripts resulting from PALB2 c.1947_1948insA visualised on 2% agarose gel. FSIs were recorded for three variant nucleotides and their corresponding wild-type nucleotides at the heterozygous positions. (d) Image of transcripts from PALB2 c.2982_2983insT visualised on 2% agarose gel. FSIs were recorded for three variant nucleotides and their corresponding wild-type nucleotides at the heterozygous positions. NCT-CT: the difference in FSI proportions in the CT and NCT conditions indicates the semi-quantitative change in relative gene expression levels of the mutant transcripts in each condition. CT, cycloheximide treated; FSI, fluorescence signal intensity; NCT, non-cycloheximide treated; NT, nucleotide; WT, wild-type.