Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Jul 15;12:63. doi: 10.1186/s13073-020-00760-3

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

PMC Copyright notice

Fig. 5 — ASXL3 is a direct target of BET inhibitors and predicts drug sensitivity. a The log2 fold change of nearby gene expression in ASXL3-depleted cells, and cells treated with either DMSO, dBET6 (300 nM), JQ1 (1 μM), or IBET-151 (1 μM). b The Venn-diagram shows the overlap of downregulated genes in cells treated with dBET6, JQ1, or IBET-151 from (a). c The bar plot shows ASXL3 expression (RPM) in NCI-H1963 cells treated with different BET inhibitors, n = 2, two-tailed unpaired Student’s t test. **P < 0.01; *P < 0.05. d Real-time PCR was performed to determine the relative gene expression of EIF4E and ASXL3 in NCI-H1963 cells treated with dBET6, JQ1, or IBET-151 for 4 h or 8 h. DMSO was used as negative control, n = 3, two-tailed unpaired Student’s t test. **P < 0.01; *P < 0.05. e BRD4 ChIP-seq was performed with NCI-H1963 SCLC cells treated either DMSO or dBET6 for 2 h. The representative track shows the occupancy of BRD4 at ASXL3 locus. f Whole-cell lysates were used for western blot with BRD2, BRD3, BRD4, Pol II, CDK9, and CCNT1 antibodies in human SCLC cell line NCI-H1963 treated with dBET6 for 0, 2, 8, and 24 h, n = 3. g Whole-cell lysates were used for western blot with ASXL3, JMJD6, NSD3, and CHD4 antibodies in human SCLC cell line NCI-H1963 treated with dBET6 for 0, 2, 8, and 24 h. HSP90 was used as an internal control, n = 3. h Whole-cell lysates were used for western blot with BRD2, BRD3, BRD4, ASXL3, JMJD6, NSD3, and CHD4 antibodies in human SCLC cell line NCI-H1963 treated with different concentrations of dBET6 for 8 h; HSP90 was used as an internal control, n = 3. i 8 different human SCLC cell lines NCI-H748, NCI-H1882, NCI-H1963, NCI-H209, NCI-H1436, NCI-H1105, NCI-H889, and NCI-H2171 were treated with different concentrations of dBET6 for 72 h. The cell viability was determined by CellTiter-Glo Luminescent Cell Viability Assay, n = 6. j The protein levels of ASXL3 and BRD4 from the previously cell lines mentioned (i) were determined by western blot, n = 3. k The representative track shows H3K27Ac and H3K4me1 levels at ASXL3 locus in ASXL3-high (NCI-H1963) and ASXL3-low (NCI-H2171) SCLC cells. l The Venn-diagram shows SE-associated genes in ASXL3-high and ASXL3-low cells from (k)