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. 2022 Apr 19;12:6445. doi: 10.1038/s41598-022-09497-y

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© The Author(s) 2022

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 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/.

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Knockdown of USP32 inhibits cell growth. (A) The mRNA level of USP32 was reduced by siUSP32-2261 and siUSP32-386. Data are represented as mean ± SD of three biological replicates. (B) Left panel: Representative images of three independent western bolting analyses showing the knockdown efficiency of siUSP32-2261 and siUSP32-386. Right panel: statistical quantification of left panel. (C) MTT assay determining the cell viability of U-87 MG and U-251 MG. Data are represented as mean ± SD of six biological replicates. (D) Representative images of three independent colony formation assays showing that USP32 knockdown inhibited cell proliferation. (E) Statistical quantification of (D). (F) USP32 knockdown reduced the number of EdU⁺ cells. Bar: 10 μm. (G) Statistical quantification of (F). Data are represented as mean ± SD of three technical replicates. (H) Representative images of three independent cell cycle assays by flow cytometry. (I) Histogram showing the percentage of each cell-cycle phase in (H). One way ANOVA followed by Tukey’s post-hoc test: vs siRNA NC, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.