MiR-29 and Mcl-1 protein expression in primary AML samples with monosomy 7. (A) This panel shows the correlation between miR-29b expression by microarrays and Mcl-1 expression by Western blotting. (Top) The microarray normalized expression of miR-29b (log2 values) for 9 patients with AML (samples 1-9). Samples 6, 8, and 9 are primary AML blasts with −7 in the context of complex karyotype (> 3 cytogenetic abnormalities), whereas the others have the following karyotypes: complex karyotype (samples 1 and 4), t(9;11) (sample 2), normal karyotype (FLT-3-ITD+, NPM1 wt) (sample 3), isolated −5 (sample 5), and normal karyotype (FLT3-ITD-, NPM1 wt) (sample 7). The miR-29a microarray (log2) value for the same 1 to 9 patient samples is as follows: 8.23, 7.76. 7.36, 7.67, 8.31, 6.5, 8.1, 6.7, and 5.7. Immediately below there is the Mcl-1 protein expression by Western blotting. K562 total cell extracts were used as positive controls for Mcl-1 expression. This cell line has undetectable miR-29b expression level. GAPDH was used as a loading control for the Western blotting. (B) Chromosomal structure of the miR-29 gene family. The diagram illustrates the position and orientation of miR-29b-1 and miR-29a cluster located on human chromosome 7, and miR-29b-2 and miR-29c cluster on human chromosome 1. Note that both clusters are located in the antisense strand. (C) MiR-29a, -29b, or -29c expression by quantitative RT-PCR in 43 primary AML samples with monosomy 7 (−7) or other cytogenetic abnormalities (Other CG). The results are shown as miRNA expression after normalization with 18s and 2ΔCt calculations. Bars represent the mean.
