Figure 1

Oncogenic HRas induces differentially expressed genes in neutrophils and transformed cells in zebrafish larvae. (A) Schematic of Translating Ribosomal Affinity Purification (TRAP) procedure. Wild-type embryos were injected with the TRAP plasmid (pTol2-Krt4-EGFP-L10a) and either control (pTol2-Krt4-HRas-mcherry) or oncogenic transformation (pTol2-Krt4-HRas^G12V-mcherry) constructs. Immunoprecipitation and sequencing were conducted 3dpf on batches of ~50 larvae collected on three separate days (n=3). Tg(LyzC : EGFP-L10a) or Tg(mpeg:EGFP-L10a) larvae were injected with either control (pTol2-Krt4-HRas-mcherry) or oncogenic Ras (pTol2-Krt4-HRas^G12V-mcherry) constructs for neutrophil- or macrophage-targeted L10a expression, respectively. (B) Cell type specific genes expressed in keratinocytes (krt17, krt15, krt97, krt91, krt8, krt92, krt18, krt94), neutrophils (mpx,lyz), or macrophages (mpeg1.1, irf8, csf1rb, spi1a, mfap4, irg1, csf1ra, spi1b, ncf1, cyba) and their expression profiles in TRAP samples from keratinocytes, neutrophils, and macrophages. (C) Venn diagram showing distribution of genes altered >2fold by TRAP-seq in response to HRas^G12V expressing keratinocytes (n=3).