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. 2022 Feb 23;13:898. doi: 10.1038/s41467-022-28566-4

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© The Author(s) 2022, corrected publication 2022

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. 4 — a, b Impact of LZTR1 loss on a RAS-GTP activity as measured by RAS-GTPase activation ELISA assay, and b RAS levels, 5 days after shLZTR1 infection. Data in a are expressed as the average of triplicate or duplicate wells with error bars indicating 95% confidence intervals. LZTR1 knockdown showed a statistically significant increase of RAS-GTP activity (P = 0.006; two-sided, unpaired Wilcoxon rank-sum test). c Effect of LZTR1 loss on MAPK activity. d YUSIK melanoma cells were incubated with the PLX4032 (500 nM) or LY3009120 (100 nM), BRAF, and pan-RAF kinase inhibitors, respectively, for 4 h at the end of treatment with LZTR1 shRNA. e RAS translocation to the cytoplasm in response to shLZTR1. RAS is visualized by staining with magenta, and GM130 or calnexin with green (Cy2). Scale bar = 50 µm. shRNAs are indicated in panels a–d by numeric identifiers; C scrambled shRNA control. Actin levels in a–d show protein loading. Cell lines indicated above the plots are colored according to their origin: acral melanoma (blue), sun-exposed melanoma (red), and normal melanocytes (gray). NBMEL newborn melanocytes. Panels a, b, d, and e represent a single experiment; panel c represents one of two experiments. Source data are provided as a Source Data file.