ZNF423 is transcriptionally up-regulated upon BMP signaling. (A) High resolution genomic profiling of ZNF423 and B cell differentiation related factors in ZNF423⁺ ALL. For indicated ZNF423 expression refer to Fig.1 B. Values were normalized to β-2-microglobulin, B2M (2^−ΔCt*1000). Error bars represent SD of technical triplicates. SNP array analysis of n = 20 initial B-ALL samples and n = 20 matched MNCs from remission BM samples. Smoothed copy-number SLR exhibiting gains or losses in at least one sample were chosen. For genes marked by *, probes located in a window of ±5 kb up- and downstream of the genomic locus are included due to scarce coverage of SNPs in coding regions. (B) Western blot of SMAD1 and phosphoSMAD1/5. BMP2-dependent SMAD1/5 phosphorylation in SEM cells treated with 100 ng/ml BMP2 1 h before lysis versus solvent treated cells as control. Actin was used as loading control. Blot is representative of two experiments. (C and D) RT-PCR analysis of ZNF423 and RUNX2 after stimulation with BMP family members. SEM cells were treated with 100 ng/ml BMP2 for indicated hours (C) or for 6 h with 100 ng/ml of BMP4 and BMP7 (D). Relative fold induction was measured by qPCR (2^−ΔΔCt), normalized to B2M and solvent control. RUNX2 expression was used as a positive control. Error bars represent SD of technical triplicates. Significance is calculated using 2^−ΔCt values by Student’s t test (*, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; n.s., not significant). Data were reproduced in three independent experiments.