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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. 3 — a Total intracellular iron in U87MG, U251 and P3#GBM glioma cells after transfection with si-COPZ1#1 at 24 h. b Graphic representation of the levels of intracellular ferrous iron compared to ferric iron, as evaluated with the colorimetric iron assay kit. c Representative fluorescence images of U87MG and U251 cells stained with the JC-1 probe to assess mitochondrial membrane potential. Scale bar, 25 μm. d Statistical analysis of the red and green fluorescence shown in (c). e Images from transmission electron microscopy showing morphology of mitochondria (white arrows) and formation of autophagosomes (red arrows) in U87MG cells transfected with si-COPZ1#1 for 24 h. Mitochondria show increased membrane density (white arrows) and a shrunken morphology. Scale bar,1.2 μm (left) and 0.6 μm (right). f Western blot analysis of TFR, TF, and FTH1 in lysates prepared from U87MG, U251, and P3#GBM cells transfected with si-COPZ1#1. Student’s t test: *p < 0.05, **p < 0.01, ***p < 0.001.