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. 2021 Sep 15;11:18345. doi: 10.1038/s41598-021-97869-1

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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 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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Fluorescent staining of CSE-treated, SCAL1-overexpressing and SCAL1-deficient A549 cells with Phalloidin-FITC and Hoechst 33342 (Olympus IX3-HOS, ×400, FITC: 500 ms). (a) A549 cells exposed to different CSE dilutions showed formation of structures associated with migration and invasion such as invadopodia (red arrowheads), hair-like filopodia (orange arrowheads), thick peripheral actin (white arrows), tunneling nanotube (TNT)-like structures (fuchsia arrows), multilobulated nuclei (yellow arrows), multinucleated cells (white brackets), and lamellipodia (orange brackets). (b) Empty vector-transfected control cells showed a stable cytoskeleton with prominent radial protrusions/focal adhesions (yellow arrowheads). Cells transfected with SCAL1 displayed transient structures such as circular dorsal ruffles (thick yellow orange arrows), peripheral dorsal ruffles (yellow brackets), stretched nuclei (cyan arrows) and perinuclear actin rims (fuchsia brackets). (c) A549 cells transfected with SCAL1-siRNA were more detached from one another and exhibited a more organized cytoskeleton with prominent stress fibers compared to siNeg controls.