Figure 5.

KHSRP is involved in regulating cell cycle and mitosis, as well as influencing the protumorigenic extracellular environment in vitro. A: Cell cycle analysis of SW480 cells transfected with siRNAs targeting KHSRP (siKHSRP) or Scramble negative control siRNA; a representative histogram of propidium iodide staining from one experiment is shown, along with a bar graph of cell distribution into cell cycle phases across three replicate experiments. An analysis of variance was performed. B: Analysis of mitotic cells by quantitation of phosphorylated histone H3 in SW480 cells transfected as in A; a representative histogram is shown with bar graph of three replicate experiments. The boxed areas indicate mitotic cells (positive for phospho histone H3 and with 2X DNA content). A Mann-Whitney test was performed. C: Wound-healing assay of SW480 cells transfected as in A; representative images from one experiment are shown, with quantitation of wound area from two replicate experiments. D: Invasion of SW620 cells transfected with siKHSRP or Scramble negative control siRNA, measured by the capacity to migrate through Matrigel; quantification of invaded cells from three replicate experiments is shown, along with representative images. E: Endothelial tube formation of human dermal microendothelial cells (HDECs) treated with conditioned media (CM) from SW620 cells transfected as in D. Tube formation is expressed as percentage of control (HDECs treated with the same media without previous conditioning in SW620 cells). A t-test was performed. F: Expression of secreted IL-8 and vascular endothelial growth factor (VEGF) in the CM of SW620 cells transfected as in D, measured by enzyme-linked immunosorbent assay. A t-test was performed. *P < 0.05, **P < 0.01. Original magnification, ×10 (C–F). AU, arbitrary unit.