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. Author manuscript; available in PMC: 2016 Jan 23.
Published in final edited form as: Oncogene. 2014 Sep 29;34(30):3935–3945. doi: 10.1038/onc.2014.324

Figure 3. AIB1 enhances Notch signaling.

Figure 3

(a, b) Knockdown AIB1 significantly decreases Hes1 promoter activity in RKO and CT26 cells as determined by luciferase reporter assay. (c) AIB1 cooperated with NICD and MAML1 to enhance Hes1 promoter activity. (d) Schematic of the Hes1 proximal promoter. Forward and Reverse: Forward and Reverse primers used for ChIP assay. NREs: Notch response elements. TSS: Transcriptional start site. (e) AIB1 was recruited to the Hes1 proximal promoter in the absence and presence of ectopic NICD as measured by ChIP assay. (f) Simultaneous recruitment of AIB1 and NICD to the Hes1 proximal promoter was measured by ChIP-reChIP assay. Cells were transfected with HA-NICD and Flag-AIB1 expression plasmids and then lysed for ChIP and ChIP-reChIP assays using control IgG, anti-HA antibody, and anti-Flag antibody for IP. (g) NICD rescued the proliferation of AIB1-knockdown CT26 cells. Each experiment was performed at least twice with similar results. All data are the means + s.d. (n=3) at each time point. Statistically significant difference: *P<0.05 and **P<0.01(t-test).