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. 2014 May 30;5(13):4821–4833. doi: 10.18632/oncotarget.2043

Figure 2. Restoring miR-31 expression inhibits DAOY cell growth, colony formation and xenograft tumorigenesis.

Figure 2

(A) DAOY cells were transfected with constitutive miR-31 expression vector, and subsequently selected in puromycin. Monoclonal cell clones were collected and subjected to stem-loop RT-PCR analysis. DAOY cells carrying the empty MSCV vector were used as the control. (B) MiR-31 expressing DAOY cells and vector control cells were seeded onto a 96-well plate. Cell viability was assessed in six consecutive days by MTT assay. (C) Histograms of distribution of DNA content in vector control and miR-31 expressing DAOY cells by flow cytometry. The percentages of cells in G1, S and G2 phase of the cell cycle were determined by analysis with the Multicycle computer software. (D) Results of colony formation assay are presented. 500 cells seeded onto a p60 plate and allowed to grow until visible colonies appeared. Colonies were stained with Giemsa. The representative figure from each group photographed (on left panel). Statistical analysis of the colony number from each group was shown on the right panel. (E) Nude mice were injected subcutaneously with vector control and miR-31 expressing DAOY cells as described in the Material and Methods section. The representative figure of mice from each group photographed at time of sacrifice (on left panel). Statistical analysis for tumor volumes from control (MSCV) and miR-31 expressing group (P-miR-31) at different time point was shown on right panel.