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. 2019 Sep 2;10:3914. doi: 10.1038/s41467-019-11884-5

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© The Author(s) 2019

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. 7 — Roles of Nuclear factor I proteins in YAP1-MAMLD1-driven tumorigenesis. a, b Immunoprecipitation assay of Flag-tagged a Nfia and b Nfib and HA-tagged YAP1 fusion-related proteins in HEK293T cells. Asterisks indicate overexpressed proteins according to their predicted molecular weights. c An enriched heatmap of YAP1 and NFIA ChIP-seq signals within the regions of ST-EPN-YAP1 group-specific YAP1 peaks. d–g IHC of P0 brains with anti-GFP (green) and anti-YAP1 (red) antibodies after electroporation of YAP1-MAMLD1-IRES-Luc and either d, f EGFP alone or e, g Nfia-En-IRES-EGFP. Arrowheads in g indicate the cells expressing exogenous YAP1-MAMLD1 but Nfia-En-EGFP. f and g are high magnification views of the area outlined by a rectangle in d and e, respectively. PB Piggybac transposase. h–k IHC of P0 brains with anti-GFP (green) and anti-Ki67 (red) antibodies after electroporation of YAP1-MAMLD1-IRES-Luc and either h, j EGFP alone or I, k Nfia-En-IRES-EGFP. Arrowheads in k indicate abnormally proliferating cells lacking Nfia-En expression. j and k are high magnification views of the area outlined by a rectangle in h and i, respectively. Scale bar in i is 100 μm (for d, e, h, and i) and the scale bar in k is 25 μm (for f, g, j, and k). l The graph represents the percentages of YAP1- and Ki67-positive cells within EGFP-positive cells. Data obtained from three independent brain samples. **p < 0.0001, n.s., not significant. The error bars indicate mean ± S.D. (n = 6). m The graph indicates the percentage of YAP1- and cleaved caspase 3-positive cells within EGFP-positive cells. p = 0.554. The error bars indicate mean ± S.D. (n = 4). n The graph indicates the percentage of YAP1- and Ki67-positive cells lacking EGFP expression. **p < 0.0001. The error bars indicate mean ± S.D. (n = 6). o, p Dorsal views of the brains bearing tumors from the mice electroporated with YAP1-MAMLD1-Luc with EGFP (o) and Nfia-Rn-IRES-EGFP (p). EGFP signals are detected only in (o). Scale bar, 1 mm. q The graph shows the percentage of EGFP-positive cells in the tumor tissues. **p < 0.0001. The error bars indicate mean ± S.D. (n = 6). Significant differences for l–n, q were assessed by t-test