Figure 1. ZFX knockdown impairs hESC colony size.

Human ESCs were transduced with ZFX knockdown lentiviral constructs and a scrambled control before clonal replating. (A) Live cell images and (B) and the entire well stained with crystal violet seven days after replating. (C) ZFX immunofluorescence after knockdown in hESCs, and (D) ZFX quantitative immunofluorescence analysis. Each dot is the average pixel intensity of nuclear ZFX protein averaged from all cells in one microscopic field. The average pixel intensity from 50 microscopic fields derived from 3 independent experiments is shown. The crosshairs and whiskers represent the mean and SEM. All knockdowns were significantly different from the scrambled control (p<0.0001 for t-tests of each knockdown versus scrambled). 11,803 (Scr), 10,296 (Z2), 4,765 (Z3) and 6,905 (Z4) nuclei were measured in these experiments. (E) ZFX RNA levels as measured by quantitative PCR after knockdown.