Figure 3.

In PDA cells, E47 directed an integrated program of gene expression that controlled cell-cycle progression. (A–E) KEGG cell-cycle pathway reflecting E47-dependent alterations in expression of cell-cycle control genes in each cell line based on RNA-seq data. (F) Western blot analysis after induction of E47 showed increased CDKN1A/p21^CIP1 protein. (G) Western blot analysis after induction of E47 showed increased CDKN1B/p27^KIP1. (H) shRNA knockdown of CDKN1B/p27^KIP1 in PANC-1/E47 cells resulted in an increased number of Ki67-positive cells after induction of nuclear E47. Data represent the mean of 6 separate fields, each with a minimum of 100 cells analyzed by an unpaired t test. Error bars are ± SEM. (I) Left: Western blot analysis showed increased p53-pSER15 in response to 2 × 1610 cGy of γ-irradiation (20 min of exposure on 2 consecutive days using a Gammacell40 Exactor, Research Irradiator with cesium 137 radiation source) to induce the DDR. Cells were harvested 3 hours after the second exposure. Right: Western blot analysis showed no increase in p53-pSER15 in response to induction of E47. ∗∗P ≤ .01. ∗∗∗∗P ≤ .0001.