Figure 2.

The HER2 c.1899‐1G>A variant resulted in aberrant splicing of exon 16. (A): Minigene assay was performed using human embryonic kidney 293T cells to investigate the impact of HER2 c.1899‐1G>A on the splicing of exon 16. HER2 exon 16 coding sequence are indicated by black boxes, 150 base‐pairs of 5’ and 3’ flanking intronic sequences are indicated by dash lines, two exons derived from SERPING1/CINH gene are indicated by dark gray boxes, and polymerase chain reaction (PCR) primers are indicated by black arrowhead. (B): Reverse transcription PCR (RT‐PCR) analysis of the spliced transcripts expressed from the wild‐type and mutant (c.1899‐1G>A) minigene constructs. RT‐PCR products were analyzed by agarose gel separation followed by sequencing of the different bands. The inclusion or the exclusion of HER2 exon 16 in each transcript is schematically indicated on the right. Abbreviations: F, forward primer; R, reverse primer; VAR, mutant construct containing the c.1899‐1G>A variant; WT, wild‐type minigene construct containing reference sequence.