Figure 2

Identification of BF104016-NSMCE2 in HL60. (a) Representative SKY karyotype of HL60 cells. Arrowheads indicate material inserted from chromosome 8 on ins(2;8) and der(13)hsr(8). Inset shows pseudocolor image of ins(2;8). (b) Copy number changes at 8q24 detected by high-resolution oligonucleotide array. NSMCE2, TRIB1, MYC, PVT1 and CCDC26 are amplified in HL60. The direction of the arrows reflects the direction of the gene transcription. NG: no gene. (c) Bubble PCR products detected by nested PCR using NVAMP1 and NSMCE2-Ex7AS for the first PCR, and NVAMP2 and NSM695 for the second. M: size marker. (d) Sequence analysis of NSMCE2 fusion transcript of HL60. (e) Detection of BF104016-NSMCE2 fusion transcripts by RT-PCR. Primers were BF104-1S and NSMCE2-Ex7AS for 5'-BF104016-NSMCE2-3'. Lanes N: water. (Additional file 4: Table S2) (f) FISH finding of HL60 using CCDC26 probe. Intense co-localized red and green signal indicates amplification of the CCDC26 gene on der(13)hsr(8) (arrowhead). A co-localized red and green signal is seen on ins(2;8) (arrow). Multiple green signals indicate amplification of the 5’CCDC26 gene on dmins. Co-localized red and green signals show normal CCDC26. (g) FISH finding of HL60 using NSMCE2 probe. Intense green signals indicate amplification of 3’NSMCE2 on der(13)hsr(8) (arrowhead) and ins(2;8) (arrow). Multiple red and green signals indicate amplification of the NSMCE2 gene on dmins. Co-localized red and green signals indicate normal chromosomal NSMCE2. (Additional file 5: Figure S3) (h and i) DAPI pictures of metaphase cells corresponding to (f) and (g). Arrow and arrowhead indicate ins(2;8) and der(13)hsr(8), respectively.