Fig. 5: Kinetics of suntag-nanos translation.
From: Direct observation of translational activation by a ribonucleoprotein granule
a, Example image of the posterior of an embryo expressing Vasa–mApple (blue), nanos-suntag-SREmut (suntag smFISH, magenta). SunTag is detected by anti-GCN4 (green). An image of the anti-GCN4 channel is shown on the right with germplasm and soma outlined. b, Translating fractions in embryos from flies expressing transgenic suntag-nanos-WT (soma n = 6, germplasm n = 7), suntag-nanos-SREmut (soma and germplasm n = 7) or suntag-nanos-tub 3′ UTR (soma n = 11, germplasm n = 7). The data are the mean ± s.d. Pairwise statistical comparisons were conducted using two-tailed Welch’s t-test. c, Left: the posterior of an embryo expressing Vasa–-mCherry (magenta) and eIF4G–GFP (green), and zoomed images of germplasm (right), showing the enrichment of eIF4G to germ granules. d, Left: the posterior of an embryo expressing Vasa–-mCherry (magenta) and yellow fluorescent protein (YFP)-tagged PABP (green), and zoomed images of germplasm (right), showing the association of PABP puncta with germ granules. e, The intensities of polysomes (anti-GCN4 staining) in soma and germplasm of embryos from flies expressing UAS-suntag-nanos-SREmut. Quantification results from four embryos were plotted in a super-plot. Individual dots represent the intensities of individual polysomes, each colour-coded by the embryo. Each coloured circle represents the mean intensity of each embryo. The black lines and error bars are the mean ± s.d. of the four embryos. Statistical comparison was performed on the mean intensities of individual embryos using two-tailed Welch’s t-test. f, The intensities of polysomes over time during live imaging suntag-nanos mRNA translation with (red curve, mean ± s.d. of 10 curves from 3 embryos) or without (blue curve, mean ± s.d. of 23 curves from 5 embryos) photo-bleaching when time is 30 s. The elongation rate calculated from the plot is indicated. g, Polysome intensities of suntag-nanos-SREmut mRNA in germplasm (blue curve, mean ± s.d. of 35 curves from 5 embryos) and soma (red curve, mean ± s.d. of 19 curves from 6 embryos) over time with photo-bleaching when time is 30 s. The elongation rates calculated from the plot are indicated. Note that elongation rates are not notably different between wild-type and SREmut RNA. h, Representative time-lapse image of FRAP of two translation sites (arrowheads). Blue, Vasa; green, scFv–GFP. Scale bar, 500 nm.
