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. 2021 Jan 8;40(8):1425–1439. doi: 10.1038/s41388-020-01622-3

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© 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/.

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Fig. 2 — a qRT-PCR to detect COPZ1 mRNA levels in U87MG, U251, and P3#GBM cells transfected with two independent COPZ1 siRNAs, si-COPZ1#1, and si-COPZ1#2. b Western blot analysis of COPZ1 protein levels in U87MG, U251, and P3#GBM cells transfected with si-COPZ1#1 and si-COPZ1#2. c Growth curves for si-COPZ1 transfected U87MG, U251, and P3#GBM cells generated with OD 450 readings plotted over time using the CCK8 assay. d Fluorescence images of EdU assays performed on U87MG and U251 cells transfected with si-COPZ1#1. Nuclei were stained with DAPI (blue). Scale bar, 100 μm. e Graphic representation of the ratios of EdU positive cells in U87MG and U251 cells transfected with si-COPZ1#1. f Representative images of colony forming assays for U251 and P3#GBM cells transfected with si-COPZ1#1 to evaluate cell proliferation. Cells were fixed and stained with crystal violet, and colonies were counted. g Graphic representation of the number of colonies shown in (f). h LDH release assay for si-COPZ1#1 transfected U87MG, U251, and P3#GBM cells compared to their respective control cells. i Representative images of live (green)/dead (red) assays for U251 and P3#GBM cells transfected with si-COPZ1#1. Scale bar, 100 μm. j Graphic representation of the rate of dead (red) cells in U251 and P3#GBM cells transfected with si-COPZ1#1. Student’s t test for two-group comparison: *p < 0.05, **p < 0.01, ***p < 0.001; one-way ANOVA for multi-group comparisons: *p < 0.05, **p < 0.01, ***p < 0.001.