Fig. 1: Sleeping Beauty (SB) mutagenesis screens on Rb-deficient background identify overlapping oncogenic alterations in mammary and lung tumors.

a Setup of SB screens using MMTV-Cre:Rb^f/f:T2/Onc3a:R26-lsl-SB11 and MMTV-Cre:Rb^f/f:T2/Onc3b:R26-lsl-SB11 mice to identify primary mammary tumors and lung metastases driven by SB transposons on Rb-deficient background. b Macroscopic lung tumors detected by stereomicroscope were dissected and subjected to ligation-mediated PCR and deep sequencing. c PCR-based analysis of primary tumors to identify the 283 bp Rb-floxed allele (lanes 1–3) and 260 Rb deletion product following Cre-mediated recombination (lanes 1 and 3, but not 2). Source data are provided as a Source data file. Representative histology (d) and pie distribution (e) of primary and lung lesions. Pleomorphic/Squamous cell carcinoma (P/SCC), Adenosquamous carcinoma (ASC), Papillary/ Micropapillary (P/MP), poorly differentiation adenocarcinoma (PDA), Cribriform (CF). Scale bar, 100 μm. f Top gene-centered Common Integration Sites (gCIS) identified in primary (top) and lung (bottom) tumors following SB screens on Rb-deficient background. All gCISs are listed in Supplementary Fig. 1A and shown in Supplementary Data 1 and 2. S-specific gCIS are highlighted in red, and percentage in primary and lung lesions is tabulated. Representative gCIS functionally analyzed herein are marked in green (top – Fbxw7) and blue (bottom – Wdr33, Srgap2, Fbxw4, Pten, Cdc42bpa, and Mtmr3). g Venn diagram for significant gCISs identified in primary and lung lesions and the 7 S-drivers. Arrows point to direction of transposons. h Schematic structure of the NF1 gene locus and relative location of CIS in primary tumors and lung metastasis. > denotes SB integration in the 5′ to 3′ direction of the gene; <denotes reverse direction.