Figure 1.

A) Schematic representation of the location of the MYB chromosome 6q23 and the NFIB on 9p22-23 and the BAC clones used for the FISH analysis. RP11-104D9 was used as a probe for MYB and probes RP11-79B and RP11-54D21 were used for the NFIB gene in the FISH analysis. B) Schematic structure of the MYB and the NFIB genes and the primers used for the RT-PCR and the 3′RACE analysis. The exon numbers of MYB are based on NCBI database (accession number NM_001130173). NFIB exon numbers were obtained from accession number ENSG0000147862 for NFIB in the Ensembl database; note MYB exon 10 is not included any MYB-NFIB chimeric transcripts and intact MYB. ‡MYB exon 10 is also known well as exon 9B (accession number HSU22376).

C) RT-PCR analysis of MYB-NFIB fusion transcripts using new primer sets. Asterisk points to case #161B4 where gene fusion was detected by FISH without transcript formation. D) FISH analysis using BAC clones of MYB (green) and NFIB (red) genes in transcript negative gene fusion positive ACC (394D7 case). White arrows point to the yellow signal representing the MYB and NFIB gene fusion. E) Sequence illustration shows fusion of MYB exon 15 with NFIB 3′UTR, as detected by 3′RACE in 394D7 case. F) Represents the MYB transcript expression of the 30 new cases. The red bars denote the MYB-NFIB transcript positive samples, whereas the blue bars represent the expression level in fusion negative tumors. The asterisks point to tumors with MYB/NFIB gene fusion by FISH only. Results are represented as fold increase relative to MYB expression in pooled normal salivary gland tissue.