Additional file 1: Table S1. of 5’UTR point substitutions and N-terminal truncating mutations of ANKRD26 in acute myeloid leukemia

Main clinical and laboratory features of the AML patients with ANKRD26 mutations identified by the present investigation. Table S2. Prediction of translation start codon presence in the ANKRD26 coding sequence and relative protein size. Figure S1. The stability of ANKRD26 mutant proteins is similar to that of the WT counterpart. HeLa cells were transfected by ANKRD26-FLAG WT or mutant constructs and cultured in a 12-well plate. Protein synthesis was blocked 24 h after transfection by addition to the cell culture of cycloheximide 100 mM diluted in DMSO. Control conditions were carried out by adding the same amount of DMSO alone. Cells were then lysed just before the addition of cycloheximide or DMSO (time 0) and 8, 24, and 48 h after the addition of cycloheximide or DMSO and analyzed by immunoblotting with anti-FLAG and anti-tubulin antibodies. The histograms show the amount of proteins expressed as FLAG/tubulin ratio and referred to time 0 of each condition. After the addition of cycloheximide, WT ANKRD26 expression decreased to about 60% at 8 h, to 45% at 24 h, and to 20% at 48 h. The expression was significantly lower after cycloheximide treatment compared with DMSO alone at each time point, indicating that protein synthesis was efficiently blocked by cycloheximide (***P < 0.001; **P < 0.01; *P < 0.05). Overall, mutant and WT proteins showed a similar kinetic of reduction after cycloheximide treatment. Data reported represent the mean of three independent experiments and are reported as mean ± S.E.M. Statistical analysis was performed by Student t test. Methods. (DOCX 200 kb

Odds ratio to have aminotransferase elevations in <i>MYH9</i>-RD patients as compared with the three control populations.

<p>Data represent ORs (95% CIs).</p><p>N.A. = not available</p><p>Note:</p>1<p> = Calculation of the OR is not possible since none of the 77 analysed ITP patients presented an elevation of AST.</p

Genetic analysis of the 75 <i>MYH9</i>-RD patients described in this study.

<p>Abbreviations: Ref. = references.</p

Demographic characteristics of the study populations.

<p>Demographic characteristics of the study populations.</p

Prevalence of liver enzyme alterations in the analyzed patient populations.

<p>Data are presented as n° of patients with altered value/total n° of evaluable patients (%).</p><p>N.A. = not available.</p

Immunohistochemistry for NMMHC-IIA in the liver biopsy from one patient with MYH9-RD.

<p>Liver biopsy from a 10-years-old patient with MYH9-RD caused by the p.T1155A mutation of MYH9 with persistently elevated AST, ALT and GGT. (A, B): Immunohistochemistry for NMMHC-IIA showed a signal (brown, horseradish peroxidase staining) concentrated close to the hepatocytes' plasma membrane. The distribution of NMMHC-IIA was not significantly different from that from a healthy control (C, D). Specimens were counterstained with Meyer's haematoxylin. Scale bars correspond to 10 µm.</p

Frequency of abnormal liver tests depending on the location of the MYH9 gene mutation.

<p>Mutations involving the motor domain and tale domain regions of the gene are indicated. No significant differences in the distribution of liver test alteration between the two locations of the gene defect were evident.</p

Basic clinical features of the 75 <i>MYH9</i>-RD patients described in this study.

<p>Proteinuric nephropathy, sensorineural hearing loss, and cataracts were evaluated as previously reported (Pecci <i>et al</i>, Hum Mutat 2008, reference 2).</p

Distribution of liver test levels in the four populations studied.

<p>Data for ALT, AST and GGT distributions are shown.</p

Results of follow-up of 29 <i>MYH9</i>-RD patients with alterations of liver enzymes.

<p>Results of follow-up of 29 <i>MYH9</i>-RD patients with alterations of liver enzymes.</p