Table 1.
Summary of clinical data and A20 mutations, gene copy numbers and expression of the analysed MM patients.
Fig 1.
Genetic aberrations of A20 in multiple myeloma.
a: Electropherogram of rs368271377 in exon 7: Arrows indicate the single base pair substitution. b: Electropherogram of rs143002189 in exon 9: The arrow indicates the single base pair substitution. c: Gene copy number analysis of A20 of selected cases: For the gene copy number assays two technical replicates of each samples were used. The blue bar represents the data for exon 4 and the red one for exon 6. Each bar represents the mean values of expression levels ± standard deviation (SD). Cut off for deletion—depicted as red line—was set at 0.7 through the fact that samples exhibited up to 40% non-neoplastic surrounding tissue. d: Representative immunohistochemical A20 staining of multiple myeloma samples. i and ii: multiple myeloma samples with reduced A20 gene copy number. iii and iv: multiple myeloma samples with normal A20 gene copy number.
Fig 2.
Structural analysis of A20 with and without rs143002189.
a) The methionine (Met) residue at position 788 in the amino acid chain of the zinc finger domain 7 (ZnF7) has been changed to isoleucine. b) Shows the different charge density distributions of the unmutated- (I) and the mutated ZnF7 (II). Surfaces with constant density values: 0.085 e/a03 are shown. III and IV show the corresponding spatial distribution of the electrostatic potential surrounding the molecule. Positive values are red encoded, negative values green. The surface shows constant values of the electrostatic potential with: 0.055 e/a0.
Fig 3.
mRNA expression analysis of A20 and 7 NF-κB target genes (BCL2, Cyclin D1, CCR7, CD44, CXCR2, cFlip, IRF4) of MM cases with (n = 6) and without (n = 14) monoallelic A20 deletions and of non-neoplastic bone marrow biopsies (BM; n = 6).
mRNA expression levels were calculated as relative expression in comparison with peripheral mononucleated cells serving as a calibrator. Each bar represents the mean values of expression levels ± standard deviation (SD). The comparison of the expression levels was performed by using the Mann-Whitney U test; all significant associations were corrected for multiple testing by applying a Bonferroni correction.