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Fig. 2 Gal80 RNA temporarily inhibits Gal4 activity in a dose-dependant manner.
(A) Embryos resulting from a cross of Tg[UAS:mem-TdTomato] fish were injected with a DNA encoding full-length Gal4 under the control of the β-actin promoter alone (a–c) or with 50 pg (d–f, N = 21), 100 pg (g–i, N = 23) or 200 pg (j–l, N = 20) of mRNA encoding full-length Gal80. Representative specimens are depicted at 3, 4 and 6 dpf. (Scale bars: 300 μm). (B) Red fluorescence quantification of tdTomato expressing cells. (a–c) Maximal projection of muscle fibers from a embryo resulting from a cross of Tg[UAS:mem-TdTomato] fish injected with the β-actin:Gal4 construct alone. The white arrowheads indicate the quantified cell. (d–f) Maximal projection of muscle fibers from a embryo resulting from a cross of Tg[UAS:mem-TdTomato] fish injected with the β-actin:Gal4 construct and 200 pg of mRNA encoding full-length Gal80. Z-stacks have been captured at the same settings as in a–c. (g–i) Same as d-f with over-exposure. (Scale bars: 20 μm) (h) Quantitative values of fluorescence per μm2. Each value corresponds to the average of the mean values obtained for three different regions of each cell. (C) Anti-tdTomato western blotting of 3dpf fish resulting from a cross of Tg[UAS:mem-TdTomato] fish injected with the β-actin:Gal4 construct alone or with 50, 100 or 200 pg of mRNA encoding full-length Gal80.