Figure 3

Knockdown of MONC reduces proliferation and viability of AMKL cells. A) Number of shRNA- or ctrl-transduced CMK and Meg-01 cells (n = 3). B) Fraction of Cerulean⁺ shRNA-transduced cells at indicated time points of culture is shown in relation to the ctrl construct (n = 3; Two-way ANOVA was performed to compare the mean of each construct at each time point to ctrl). C) Number of colonies from methylcellulose-based CFU assays of indicated shRNA-transduced CMK and Meg-01 cells (n = 3). D) Cumulative number of CFUs after one round of replating of sh-MONC in CMK and Meg-01 cells (n = 3). E) Percentage of shRNA-transduced Meg-01 cells in subG1 (BrdU⁻/7-AAD⁻), G1 (BrdU^low/7-AAD^low/high), S-phase (BrdU⁺/7-AAD^low/high) and G2/M fraction (BrdU^low/7-AAD^high) (n = 3). F) Percentage of apoptotic (Annexin V⁺/7-AAD⁻) and dead (7-AAD⁺) cells for shRNA-transduced CMK and Meg-01 cells measured by flow cytometry after 5 days of culture (n = 3). G-H) Merged density plots of viable, Cerulean⁺G) Meg-01 and H) CMK cells for indicated surface markers as measured by flow cytometry after 5 days of culture (n = 4); population frequencies with errors are displayed for respective framed gates. I) Number of colonies from methylcellulose-based CFU assays of indicated shRNA-transduced Down-Syndrome transient leukemia blasts (n = 2; error bars show variation). (A-H) Data are presented as mean ± s.d. *P < 0.05; **P < 0.01.