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. Author manuscript; available in PMC: 2019 Apr 15.
Published in final edited form as: Cancer Res. 2018 Jan 30;78(8):2040–2051. doi: 10.1158/0008-5472.CAN-17-2761

Figure 4. HER2-mediated sequential activation of IL1A and IL6 transcription.

Figure 4

A. Real-time RT-PCR analysis of IL1A and IL6 mRNA levels in MCF10A-HER2 cells treated with several HER2-downstream signaling pathway inhibitors for 24 hours. B and E. Schematic depiction of the potential transcriptional binding sites on the IL1A (B) and IL6 (E) promoter. C and F. MCF10A-vector and MCF10A-HER2 cells were co-transfected with IL1A (C) or IL6 (F) promoter reporter and several transcriptional factors-expression plasmids. At 48hrs post-transfection, luciferase assays were performed. RLU, relative luciferase unit; Vector, MCF10A-vector; HER2, MCF10A-HER2 cells. D and G. ChIP analysis of the relative binding abilities of PU.1 and NF-κB to IL1A (D) or IL6 (G) promoter in MCF10A-HER2 cells treated with DMSO and PD0325901 for 24 hours. The IP experiments were repeated and all of qRT-PCR reactions were performed in triplicate. H. A proposed model for HER2-mediated sequential upregulation of IL1A and IL6 expression in the context of HER2-downstream pathways and functions. For A, C, D, and F, the experiment were repeated three times and results were shown as mean ± SD. *p < 0.05, **p < 0.01. ***p < 0.001 (unpaired Student’s t test).