American Society of Hematology

Figure 3

$Figure 3. Modulation of TLE levels affects the proliferation and survival of Kasumi-1 cells. (A) ShRNAs directed against TLE1 or TLE4 knockdown expression to approximately 45% control values and are specific for their respective target gene. (B) Knockdown of TLE1 or TLE4 increases cell-cycle progression. Flow cytometric determination of G0/G1 fraction in wild-type (WT) Kasumi-1 cells and cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. (C) Knockdown of TLE1 or TLE4 increases cyclin D1 expression. Flow cytometric analysis of percentage of cyclin D1 positive Kasumi-1 cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. (D) Knockdown of TLE1 or TLE4 increases expression of the cell proliferation marker Ki-67. Flow cytometric analysis of fluorescence intensity (FL) of Ki-67 in Kasumi-1 cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. (E) Knockdown of TLE1 or TLE4 increases cell division. Flow cytometric cell proliferation assay with DiI in Kasumi-1 cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. Shown is the percentage of bright DiI positive (DiI++) cells over time. (F) Knockdown of TLE4 leads to a proliferative advantage. Relative proportion over time of GFP positive Kasumi cells either infected with a control (Ctrl) or a specific shRNA against TLE4. (G) Expression of TLE1 or TLE4 leads to an increase in apoptosis and cell death. Percentage of apoptotic cells (annexin V+/7AAD−), dead and late apoptotic cells (annexin V+/7AAD+), and total apoptotic and dead cells in Kasumi cells infected with control empty MSCV-GFP retroviral vector (Ctrl) or retrovirus expressing TLE1 or TLE4 cDNAs. GFP positive cells were sorted before analysis. (H) Expression of TLE1 or TLE4 slows proliferation. Relative proportion over time of GFP positive Kasumi cells infected with either control empty MSCV-GFP retroviral vector (Ctrl) or retrovirus expressing TLE1 or TLE4 cDNAs. All diagrams denote results from 3 independent experiments. Results are expressed as the mean plus or minus SEM (*P < .05, significant difference vs controls).$

Modulation of TLE levels affects the proliferation and survival of Kasumi-1 cells. (A) ShRNAs directed against TLE1 or TLE4 knockdown expression to approximately 45% control values and are specific for their respective target gene. (B) Knockdown of TLE1 or TLE4 increases cell-cycle progression. Flow cytometric determination of G₀/G₁ fraction in wild-type (WT) Kasumi-1 cells and cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. (C) Knockdown of TLE1 or TLE4 increases cyclin D1 expression. Flow cytometric analysis of percentage of cyclin D1 positive Kasumi-1 cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. (D) Knockdown of TLE1 or TLE4 increases expression of the cell proliferation marker Ki-67. Flow cytometric analysis of fluorescence intensity (FL) of Ki-67 in Kasumi-1 cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. (E) Knockdown of TLE1 or TLE4 increases cell division. Flow cytometric cell proliferation assay with DiI in Kasumi-1 cells infected with control (Ctrl) shRNA and shRNA against TLE1 or TLE4. Shown is the percentage of bright DiI positive (DiI⁺⁺) cells over time. (F) Knockdown of TLE4 leads to a proliferative advantage. Relative proportion over time of GFP positive Kasumi cells either infected with a control (Ctrl) or a specific shRNA against TLE4. (G) Expression of TLE1 or TLE4 leads to an increase in apoptosis and cell death. Percentage of apoptotic cells (annexin V⁺/7AAD⁻), dead and late apoptotic cells (annexin V⁺/7AAD⁺), and total apoptotic and dead cells in Kasumi cells infected with control empty MSCV-GFP retroviral vector (Ctrl) or retrovirus expressing TLE1 or TLE4 cDNAs. GFP positive cells were sorted before analysis. (H) Expression of TLE1 or TLE4 slows proliferation. Relative proportion over time of GFP positive Kasumi cells infected with either control empty MSCV-GFP retroviral vector (Ctrl) or retrovirus expressing TLE1 or TLE4 cDNAs. All diagrams denote results from 3 independent experiments. Results are expressed as the mean plus or minus SEM (*P < .05, significant difference vs controls).

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