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. 2019 Nov 26;8(12):70. doi: 10.1038/s41389-019-0179-2

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Schematic illustration of the H3K27Ac (Histone H3acetyl K27) enrichment and PCR fragments of the human IL-6 gene promoter using the UCSC genome browser. b Chromatin immunoprecipitation (ChIP) assays were performed in HeLa229P, HeLa229/TR, HeLa229 TR/Erl, HeLa229 TR/CTL, and HeLa229 TR/CRISPR cells using antibodies against MUC1, EGFR, and H3K27Ac, respectively, to identify the occupancy on IL-6 gene promoter. The ABCB1 promoter was used as a positive control for ChIP. c HeLa229P, HeLa229/TR, HeLa229 TR/CTL, HeLa229 TR/CRISPR, HeLa229 TR/shCTL, and HeLa229 TR/shEGFR cells were transfected with pGL3-IL-6 promoters or pGL3-basic plasmids (500 ng) and pGL3-renilla plasmid (10 ng), luciferase activity was detected. d HeLa229P, HeLa229/TR, HeLa229 TR/CTL, HeLa229 TR/CRISPR, HeLa229 TR/shCTL, and HeLa229 TR/shEGFR cells were transfected with pGL3-IL-6-mutant-promoters or pGL3-basic plasmids (500 ng) and pGL3-renilla plasmid (10 ng) and subjected to luciferase activity assay. The bars in d is the same as that in c chart. e The mRNA levels (left) and production (right) of IL-6 in HeLa229 TR/shCTL and HeLa229 TR/shCREBs were measured. f The mRNA levels (left) and production (right) of IL-6 in HeLa229 TR/shCTL and HeLa229 TR/shGRβs cells were detected. Data represent mean ± SD from three independent experiments (n = 3). Differences between linked groups were evaluated by two-tailed Student's t test. *P < 0.05; **P < 0.01; ***P < 0.001; ns not significant, P parental, TR paclitaxel-resistant, TR/Erl paclitaxel-resistant/erlotinib, p+386~+557: pIL-6-promoter-+386~+557, p-645~+557: pIL-6-promoter-645~+557, p-1503~+557: pIL-6-promoter-1503~+557, WT: pGL3-IL-6-Luc-wt, AP-1: pGL3-IL-6-AP-1-mutant, NF-κB: pGL3-IL-6-NF-κB-mutant, C/EBP: pGL3-IL-6-C/EBP-mutant, CREB: pGL3-IL-6-CREB-mutant, C/EBPβ: pGL3-IL-6-C/EBPβ-mutant, GRβ: pGL3-IL-6-GRβ-mutant.