Fig. 4. Knockdown of PAK2 or CCND3 inhibits tumor cell growth.

a–g HCT116 cells were transfected with si-RNAs for negative control (si-NC), PAK2 (si-PAK2), or CCND3 (si-CCND3). At 24 h after transfection, mRNA levels of PAK2 and CCND3 were determined by qRT-PCR a. At 72 h after transfection, cell viability was determined by MTS assay b. At 48 h after transfection, cell cycle was analyzed c and the percentage of cells in G1 and G2/M phase are annotated in each column. At 48 h after transfection, apoptosis was measured by flow cytometric analysis of cells stained with Annexin V-FITC and PI d. Right, the quantified apoptotic cell population was estimated At 48 h after transfection, equal amounts of cell lysates were subjected to western blotting using the indicated antibodies e. At 24 h after transfection, cells were re-plated and performed colony formation f and soft agar assays g. All data are presented as averages of triplicate measurements with error bars representing standard deviations. ***P < 0.001