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Fig. 3
Expression of the mammalian tumor suppressor ARF in zebrafish driven by heat shock promoter. In vivo analysis of transgenic zebrafish expressing human ARF under the control of an inducible heat shock promoter, Tg (hsp70l:ARF) (hs:ARF). Schematic of the hs:ARF transgene (top left). The ARF cassette included in the transgene is a cDNA that consists of human exons 1b, 2, and 3 of CDKN2A. Representative Western blot of 3 replicates of ARF before (0 hr) and 3 and 6 hr post heat shock induction of ARF expression (top middle). Portion of fin shown for analysis of expression in vivo (top right; dashed box). Scale bar: 1 mm. Immunostaining (sagittal confocal images) for ARF in adult hs:ARF zebrafish fins at 0, 3, and 8 hr after a single, hour long, 37°C heat shock (bottom). Scale bars: 50 µm. ARF expression is maximal at 3 hr post heat shock, and it is undetectable by 8 hr post heat shock. Figure supplement 1 shows in vitro assays.