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. 2024 Nov 13;13(1):39. doi: 10.1038/s41389-024-00540-3

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 4 — A Incucyte confluency monitoring of human and mouse GBM cell models as readout for cell proliferation in response to sertraline (red), chloroquine (blue) or combined treatment (green) as compared to a DMSO control (gray). B Alvetex 3D clonogenic assays in ser/gly^high 2012.2 GBM cells and ser/gly^low 1919 GBM cells upon combined sertraline and chloroquine treatment, under both normal oxygen and hypoxic conditions. Each individual dot represents an independent experiment. One-way ANOVA with Tukey’s multiple comparison test has been performed. C Alvetex 3D clonogenic assay in GFP Ctrl, PSPH^amp, PSPH^V116I GBM 1919 cells treated with DMSO, sertraline, chloroquine, or the combination treatment in low glucose conditions. Each individual dot represents an independent experiment. One-way ANOVA with Tukey’s multiple comparison test has been performed, comparing each treatment to their respective DMSO controls in each model. D Alvetex 3D clonogenic assay and the quantification of ser/gly^high 2012.2 GBM cells comparing scrambled control cells to PSPH knockdown cells for their clonogenic capacity using two independent shRNAs, shPSPH #1 and #2 respectively (representative of 3 biological repeats). One-way ANOVA with Tukey’s multiple comparison test has been performed compared to the scrambled control. E Incucyte confluency monitoring of GBM cells as readout for cell proliferation 72 h after transduction plating 2000 cells per well in 6 replicates to profile the effect of PSPH knockdown (shPSPH1 blue, shPSPH2 light blue) in ser/gly^high 2012.2 GBM cells as compared to scrambled control cells and in response to chloroquine (striped line) treatment. One-way ANOVA with Tukey’s multiple comparison test has been performed to determine the effects of PSPH targeting compared to the scrambled control. For CQ effectiveness over time multiple unpaired t tests with False Discovery Rate (FDR) was performed with q-values < 0.05 determining the actual point of CQ impact on growth curve deviation. Statistical significance p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), and q < 0.05 (*), q < 0.001 (***).