Cloning efficiency of leukemic cell lines on lentiviral transduction with shRNA against AES
| . | Scrambled control . | shRNA-AES-#1 . | shRNA-AES#2 . |
|---|---|---|---|
| Kasumi-1 cloning | 1 in 15 | 1 in 39 | 1 in 109 |
| 95% CI | 12-17 | 33-46 | 92-129 |
| P | .0001 | .0001 | |
| KCL-22 cloning | 1 in 82 | 1 in 72 | 1 in 79 |
| 95% CI | 65-104 | 57-91 | 63-100 |
| P | .42 | .82 | |
| K562 cloning | 1 in 28 | 1 in 20 | 1 in 43 |
| 95% CI | 22-35 | 16-25 | 34-55 |
| P | .05 | .01 | |
| U937 cloning | 1 in 2 | 1 in 3 | 1 in 3 |
| 95% CI | 2-3 | 2-3 | 3-4 |
| P | .29 | .04 |
| . | Scrambled control . | shRNA-AES-#1 . | shRNA-AES#2 . |
|---|---|---|---|
| Kasumi-1 cloning | 1 in 15 | 1 in 39 | 1 in 109 |
| 95% CI | 12-17 | 33-46 | 92-129 |
| P | .0001 | .0001 | |
| KCL-22 cloning | 1 in 82 | 1 in 72 | 1 in 79 |
| 95% CI | 65-104 | 57-91 | 63-100 |
| P | .42 | .82 | |
| K562 cloning | 1 in 28 | 1 in 20 | 1 in 43 |
| 95% CI | 22-35 | 16-25 | 34-55 |
| P | .05 | .01 | |
| U937 cloning | 1 in 2 | 1 in 3 | 1 in 3 |
| 95% CI | 2-3 | 2-3 | 3-4 |
| P | .29 | .04 |
Cloning efficiency was calculated by plating 1, 3, 10, 100, and 300 transduced cells (GFP+) each into 48-well colony assays (300 cells only for Kasumi-1 and KCL-22). The number of wells with colony growth was counted. Poisson distributions were calculated using L-Calc limiting dilution analysis software Version 1.1.1 (StemSoft Company). The calculated cloning efficiency as well as the 95% confidence intervals (CI) are indicated. Two-sided P values were calculated for each cell line and each shRNA construct with regard to the cloning efficiency in the scrambled control. The data indicate the mean of 2 independent experiments. Strong effects of shRNA against AES were found only in AML1/ETO+ Kasumi-1 cells with 2.6-fold and 7.3-fold reduction in cloning efficiency. Minor effects (< 2-fold reduction) were observed in K562 cells and in U937 cells.