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. 2023 Dec 14;42:338. doi: 10.1186/s13046-023-02922-8

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© The Author(s) 2023

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Nutlin-3a induces huge cytoplasmic vacuoles in KRAS MT/p53 WT NSCLC cells. Cells were treated with nutlin-3a (30 μM) for 6 (D, H) or 24 h (A, B, E–H). A, B Representative live cell images captured by light (A) and holotomography microscope (B) show huge cytoplasmic vacuoles in KRAS MT/p53 WT but not KRAS WT cells. C Time-lapse video capture images (24 h) of H460 obtained by holotomography microscopy show increase in macropinosomes volume with time and membrane rupturing in the end. D Nutlin-3a induced formation of huge vacuoles were incorporated with dextran (green), but not merged with LysoTracker (red). Representative live images were obtained by confocal laser scanning microscopy (left). Number of macropinosomes were quantitated from the captured image (right) (n ≥ 8). E–G Representative images obtained by confocal laser scanning microscopy. Fixed cells were stained with Rab7 (E), LAMP1 (F), or Rac1 (green), and rhodamine–phalloidin (red) (G) and were counterstained with DAPI (blue). H Western blotting of Rac1 expression. *p < 0.05, **p < 0.01, ***p < 0.001 compared with control. Scale bar: 200 μm (A), 7 μm (B, C), and 20 μm (D-G)