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Fig. 1
Generation and characterization of Tg(ifabp:EGFP-krasV12) transgenic zebrafish. (A) Schematic diagram showing the DNA pDs-ifabp:LexPR-Lexop:EGFP-krasV12 construct used for the generation of Tg(ifabp:EGFP-krasV12) transgenic fish. Ds, maize Ds transposon sequence. (B) Intestine-specific expression of EGFP-krasV12 in F1 transgenic fry and wild-type fish, and confocal image of whole mounted kras+ transgenic larva at 8 dpf following induction by 1??M Mifepristone. The images were obtained at z?=?5??m over a thickness of 50??m. (C) Western blot analysis of total proteins from WT and kras+ transgenic fish in the gut tissue from three founder lineage (I, II, and III) for the detection of the Kras protein. ?-Actin, internal control for equal loading. (D) RT-qPCR analysis of kras RNA expression in the kras+ transgenic F2 whole larvae from founders I and III. N?=?20 larvae from each founder (at 2 dpi using 1??M mifepristone) were used for isolation of proteins and mRNA. (E) Fluorescence micrographs of intestine cross sections from wild-type and kras+ zebrafish larvae showing green fluorescence or phalloidin staining. (F) Quantification of villi by ratio of inner to outer circumferences of intestine at 10 dpi by 2??M mifepristone. Statistical significance: ***P?<?.001.