Figure 3

Silencing of GAS5 shortens the cell cycle length as determined by EdU incorporation assay. A. Representative micrographs of shGAS5 cells pre-treated with or without doxycycline during 72 h then incubated with EdU for 12 hrs. Scale bar: 100 μm. B. Cell cycle length was measured from a time course of EdU incorporation. Five different fields were imaged for each group in each of three independent experiments under fluorescence microscope and number of EdU positive cells was counted in approximately 1200 cells (DAPI). The graph plots the percentage of EdU positive cells vs. time in axes. The time taken for 100% of EdU positive cells was determined from the linear regression and slopes in presence or absence of doxycycline were compared p < 0.0001. C. Time in hours needed for 30, 50, 75 and 100% of cells to become EdU positive calculated according to the linear regression method.