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. 2016 Mar 21;35(39):5155–5169. doi: 10.1038/onc.2016.47

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Copyright © 2016 Macmillan Publishers Limited

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Lpd is required for EGF-induced membrane protrusion. (a) Representative kymographs of Ctrl-shRNA and Lpd-shRNA2 MTLn3 cells. A line drawn perpendicular to the cell surface is shown for each frame of a time-lapse movie to depict temporal dynamics of cell edge. X axis: time (arrow length: 20 s); Y axis: distance (arrow length: 3.1 μm). (b) Quantification of protrusion parameters from kymographic analysis of Ctrl-shRNA and Lpd-shRNA2 MTLn3 cells. Data represented as mean±s.e.m. Unpaired t-test; *P⩽0.05. (c) Micrographs showing immunofluorescence for endogenous Lpd in MTLn3 cells stimulated with 5 nm EGF (post-stimulation time is indicated). Scale bar, 10 μm. (d) Quantification of data shown in (c): mean fluorescence intensity of Lpd within a 0.66-μm zone at the lamellipodial edge is plotted as a function of time; >30 cells analyzed from at least three independent experiments. Error bars indicate s.e.m. (e) Representative micrographs from time-lapse movies of Ctrl-shRNA (control non-targeting shRNA) and Lpd-shRNA2-expressing MTLn3 cells stimulated with 5 nm EGF. Dashed line shows cell edge. Scale bar, 10 μm. (f) Quantification of membrane protrusions on Ctrl-shRNA and Lpd-shRNA-treated cells. Cell area was determined after EGF stimulation and normalized to the pretreatment cell area; >30 cells analyzed from three independent experiments. Error bars indicate s.e.m. See also Supplementary Figure S3.