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. 2018 Dec 21;9:5426. doi: 10.1038/s41467-018-07126-9

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© The Author(s) 2018

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 — Haplo-deficient Phd2 is sufficient to cooperate with BRaf^V600E to induce melanoma in mice. a–h Haplo-deficient Phd2 is sufficient to cooperate with BRAF^V600E to induce melanoma in mice. The 6-week old adult Tyr::CreER; BRaf^CA; Phd2^lox/+ mice were treated topically on the ear, flank and tail with 4-OHT. The presence of pigmented lesions was assessed at days 45 (a), 90 (b), 135 (c) and 180 (d) following 4-OHT administration. 4-OHT-treated mice were killed at 45, 90 and 180 days after the induction. The pigmented lesions were photographed at day 45 (e), 90 (f) and 180 (g, h) following the 4-OHT induction. i Kaplan–Meier survival analysis of Tyr::CreER; BRaf^V600E; Phd2^-/+ neonatal mice (n = 20) and Tyr::CreER; BRaf^V600E; Phd2^-/+ (n = 20) adult mice (6 weeks to 1 year old). j Comparison of tumor volume in mice with homozygous deletion of Phd2 or haplo-deficient Phd2 mice. Tumor volume in Tyr::CreER; BRaf^V600E; Phd2^-/+ (n = 20) mice was significantly smaller than that of Tyr::CreER; BRaf^V600E; Phd2^−/− mice (p = 0.00171). k Melanoma shown on uninduced part of body (%) in homozygous deletion or haplo-deficient Phd2 mice. Melanoma present at uninduced body part was quantified (n = 20 in each group). l Average number of positive lymph nodes in homozygous deletion or haplo-deficient Phd2 mice (n = 20 in each group). m, n CD31 immunohistochemical stain in in homozygous deletion or haplo-deficient Phd2 mice (n = 5 in each group). Bars in a–d indicate 1 mm. Bars in e–h, m, n indicate 100 μm. Arrows in m, n point to CD31+ blood vessels. *p < 0.05 (t-test) compared to Brafv600E;Phd2^−/− group in j and l; error bars indicate s.d.