NCAM suppression inhibits cell growth and enhances differentiation and cell death. (A) Measurement of proliferation after Dox-mediated induction of different shRNA clones directed against NCAM1 (shNCAM_1, _2, _3) in the indicated cell lines. Cell count was performed daily starting at day 4 of Dox treatment. Error bars represent mean of 3 measurements ± SD of mean. (B-C) Cell cycle (CC) analysis and measurement of apoptosis were performed at day 5 after Dox-mediated induction of different shRNA clones directed against NCAM1 (shNCAM_1, _2, _3) or Scr as indicated. Shown is the fraction of cells in subG1, G1, S, and G2/M phase (B) and the percentage of Annexin V⁺ cells (C). Statistic analyses were performed by the 2-tailed Student t test. Error bars represent mean of 3 experiments for each condition ± SD of mean. (D) May-Grünwald-Giemsa staining and expression analysis of CD11b at day 5 after Dox-mediated induction of shNCAM_3 or Scr shRNA. Scale bar, 10 μM; the enlarged cells in the corners are 4× enhanced. Shown are representative examples of myelomonocytic differentiation and CD11b cell surface expression (N = 3).