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

. 2021 Feb 15;12(2):183. doi: 10.1038/s41419-021-03475-4

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

© The Author(s) 2021

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/.

PMC Copyright notice

Fig. 6 — A Levels of DNA repair-related proteins in the parental HCT-15 and their GSK3β-depleted single clone (KO1 and KO2) cells determined by western blotting. B Levels of DNA repair-related proteins in HCT-15 cells following treatment with 10 μM CHIR99021 HCl (CHIR) or 5 μM LY2090314 (LY) determined for 48 h by western blotting. C BRCA1 protein level was partially restored in HCT-15 GSK3β-depleted cells transfected with full-length WT-GSK3β cDNA (WT) but not with mutated-GSK3β cDNA (Y216F). The relative intensities of the bands were quantified by Image J software and normalized to GAPDH levels. All data are expressed as mean ± SD from three independent experiments. (*p < 0.05, **p < 0.01, ***p < 0.001, t test). D Exogenous expression of GSK3β in HCT-15 cells increased BRCA1 protein level. The relative intensities of the bands were quantified by Image J software and normalized to GAPDH levels. All data are expressed as mean ± SD from three independent experiments. (*p < 0.05, t test). E mRNA expression of BRCA1 in the parental HCT-15 and their GSK3β-depleted single clone (KO1 and KO2) cells was detected by qRT-PCR. (n = 3, *p < 0.05, t test). F BRCA1 mRNA levels in HCT-15 cells treated with GSK3i, CHIR99021 HCl (CHIR), and LY2090314 (LY), for indicated times and concentrations. (n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, one-way ANOVA). G and H The Snail and Slug expression levels were negatively correlated with the levels of BRCA1. The expression of indicated proteins was analyzed by western blotting in the HCT-15 cells transfected with the full-length Wnt3a cDNA (G), or treated with GSK3i LY2090314 (LY) and depleted GSK3β (KO; H). I Knockdown of Snail and Slug increased BRCA1 expression in HCT-15 cells. Cells were treated with siRNAs targeting human Snail, Slug, or siNC for 48 h. The expression of indicated proteins was analyzed by western blotting. J Silencing of Snail and Slug restored the levels of BRCA1 in GSK3β KO cells. HCT-15 and GSK3β KO cells were treated with siSnail, siSlug, or siNC for 48 h. K and L Effect of single agent and combination treatment on indicated cells viability for combinations of PARP inhibitor, simmiparib (SP), plus GSK3 inhibitor LY2090314 (LY). UWB1.289 (carrying a BRCA1 mutation) and UWB1.289 + BRCA1 (restored with wild-type BRCA1) cells were analyzed 7 days after treatment with simmiparib (SP), LY2090314 (LY), or a combination (K). HCT-15 and RKO cells were transfected with siBRCA or siNC for 24 h and then followed by treatment of simmiparib (SP), LY2090314 (LY), or a combination for 7 days (L). Cell viability was measured by SRB assay. Combination index (CI) was calculated using CompuSyn software with the Chou–Talalay equation, and average CI values are presented (CI < 1, synergism; CI = 1, additive effect; CI > 1 antagonism). Data are from three independent experiments and expressed as mean ± SD.