Absence of BCL10 Mutations in Human Malignant Mesothelioma

Figure 1. Genetic Analysis of BCL10 in Germ Cell Tumors (GCTs) and B Cell Lymphomas (A) PCR-SSCP analysis. (a) and (c), GCT; (b) and (d), lymphoma. (a) and (b), exon 1; (c) and (d), exon 3.2. Arrows indicate conformational changes. N, normal; T, tumor; CL, cell line; tumor numbers are indicated on top. (B) PCR sequence analysis. (a) and (b), Tera-1; (c), GCT-44; (d) and (e), Tera-2; (f), T-243A; (g)–(i), 240A. Panels (a)–(e) show absence of mutations in GCT cell lines in relation to Willis et al. (1999) report, where boxes indicate base change and upward arrow indicates insertion positions. Positions in coding region of cDNA are: (a), 172 bp; (b), 653 bp; (c), 172 bp; (d), 58 bp; (e), insertion at 499 bp. Panel (f), A/G heterozygosity at position 2 of codon 213. Panels (g)–(i) indicate loss of heterozygosity at codon 5 (underlined) in tumor and cell line 240A. Panel (g), normal DNA; panel (h), tumor DNA; panel (i) cell line DNA. The residual peak of G in panel (h) indicates the presence of contaminating normal cells. (C) Multiplex RT-PCR analysis of BCL10 expression in GCT. Primers from the 59 coding region (59-GGACCCGGAAGAAGCGCCATC TCC-39 and 59-AAGTAGTCTAACAATTTTCCA GCCC-39) spanning two different exons were used to amplify a 249 bp PCR product. PCR was performed utilizing AmpliTaq DNA polymerase (PE Applied Biosystems, Foster City, CA). The 450 bp PCR product represents b-actin as control. Marker, PhiX HaeIII; tumor and cell line numbers are shown on top.