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. 2017 Dec 11;25(5):935–950. doi: 10.1038/s41418-017-0030-2

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© The Author(s) 2017

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) HMEL immortalized melanocytes were infected with GFP lentiviral particles expressing empty vector (EV), FLAG-LOXL3, or FLAG-LOXL3Δ and GFP-positive sorted cells were analyzed for the expression of LOXL3 isoforms by western blot. β-actin was used as a loading control. MW of protein standards (kDa) is indicated. (b) Soft-agar colony formation assay of HMEL melanocytes transduced as in (a). Error bars represent s.e.m., n = 4 biologically independent replicates. (c) Cell proliferation of HMEL melanocytes transduced as in a. Cell proliferation is shown for each cell line with three experimental replicates; error bars represent s.e.m. The experiment was repeated twice with similar results. (d and e) HMEL immortalized melanocytes transduced as in (a) were analyzed in migration (d) and invasion (e) assays. Graphs show the number of migrating and invading cells quantified after cell fixation and staining. Error bars represent s.d., n = 3 biologically independent experiments. (f) A375P NTC, shL3#1 and shL3#2 xenografts were grown in nude mice (n = 8 for NTC, n = 7 for shL3#1 and n = 6 for shL3#2) and tumor volume was measured every week (left). Error bars represent s.e.m. The percentage of tumors developed (incidence) in each group and the required days from cell injection until the tumors were measurable (latency) is shown on the middle. The average latency and minimal and maximal days for latency in each group is also indicated (right). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 by a two-sided Student’s t-test (b and d–f)