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. 2022 Mar 24;12:5076. doi: 10.1038/s41598-022-09078-z

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© The Author(s) 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 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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Effects of AXLi mono- and additive treatments. Ten days (twice/day) treated Melmet 1 tumors (BRAFi: 12.5 mg/kg, AXLi: 50 mg/kg, the combination or vehicle (DMSO/methylcellulose)) were subjected to RPPA analysis and AXLi-induced effects on protein expression either upon mono-treatment “AXLi vs. control” (a) or as an additive effect when combined with BRAFi “Combination vs BRAFi” (b) presented as volcano plots. Differentially expressed proteins: only p value < 0.05 (red), only log2 fold change difference (log₂FC) > 0.5 (yellow) and both p value < 0.05 and log₂FC > 0.5 (green). Not significantly (NS) changed proteins indicated in grey. Positive and negative log₂FC values represent increased and decreased protein expression, respectively, in AXLi vs Control or AXLi + BRAFi vs BRAFi treated tumors. (c) Expression levels of proteins (after normalization to the loading control, α-tubulin) measured by SWI in the treated tumors as indicated. Average ± St.Dev., n = 2. (d, e) Top three identified functional enrichments (ranked based on strength set to be above 1.5) within KEGG⁶⁵ and Reactome signaling pathway databases, analyzed by STRING (www.string-db.org), in AXLi- mono-treated (d) or as a part of combination with BRAFi (e). Strength and False Discovery Rate (FDR) describe how large and significant, respectively, the enrichment effect is.