Effects of Zfp423 expression on the colony formation and proliferation of p210BCR/ABL-expressing cells. (A) Schematic structures of the retroviruses and the illustration of the experimental procedure. BM cells were extracted from WT or p210BCR/ABL transgenic mice, infected with empty retrovirus (pMyIG, control) or Flag-HA–tagged Zfp423 (FHZfp423)–expressing retrovirus (pMyIG/FHZfp423, FHZfp423), and subjected to the B-cell colony assay. (B) Results of B-cell colony assay. The mean green colony number of 3 independent experiments for each group (WT+conrol, p210BCR/ABL+control, WT+FHZfp423, and p210BCR/ABL+FHZfp423) is shown with error bars. (C) Suppression of ZNF423 expression by shRNAs. mRNA (5 μg) extracted from the parental BV-173 line and 2 shRNA-introduced sublines (shRNA-1 and shRNA-2) were blotted with a part of the human ZNF423 coding region. β-Actin hybridization was performed as an internal control and the relative expression ratio of ZNF423 to β-actin in each cell line is shown as a vertical column. (D) Results of cell proliferation assay. Cells of the parental BV-173 line and 2 shRNA-introduced sublines (BV-173/shRNA-1 and BV-173/shRNA-2) were plated at a density of 105/10 cm2 on day 1 and cell numbers were counted on day 3 and day 5. The mean cell number of 3 independent experiments of each line is plotted with error bars.
