Fig. 1

Fig. 1 Generation and characterization of Tg(fabp10:EGFP-kras^V12) transgenic zebrafish. (A) Schematic diagram of the DNA construct used to generate Tg(fabp10:EGFP-kras^V12) transgenic zebrafish. Ds, maize Ds transposon sequence. (B–G) Liver-specific expression of EGFP-Kras^V12 in F₁ transgenic fry (B,E,F,G) as compared with the Tg(fabp10:dsRFP; elaA:EGFP) transgenic line expressing RFP in liver (Korzh et al., 2008) as normal control (C,D). (H–K) Gross observation of control fish (H) and F₁ krasV12 transgenic fish (I–K). Ages of the fish are indicated. Scale bars: 2 mm. (L) Kaplan-Meier survival curves of the Tg(fabp10:EGFP-kras^V12) fish for three groups of heterozygous transgenic zebrafish from F₁/I (n=262), F₁/II (n=311), F₂/II (n=356) and WT siblings as control (n=275). (M) Determination of endogenous and transgenic kras expression levels by qRT-PCR. Log₂ fold changes for endogenous and transgenic kras mRNAs were calculated against an internal housekeeping gene (β-actin) using the C_T method. Histological analysis was performed to confirm their neoplastic stages before qRT-PCR (H, hyperplasia; C, carcinoma; N, normal liver). Results are presented as the mean ± s.d. and each bar represents five biological replicates.