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Fig. 9. Targets of MBT-activated transcription factors are found activated within 1-2 h in the third wave. (A) Onset times from Series 2 data for published targets of transcription factors Mixer (Loose and Patient, 2004; Sinner et al., 2006) and Brachyury (Gentsch et al., 2013). Numbers of targets shown at each time point: light blue, onset in second wave; dark blue, onset in third wave or beyond. Onset time of transcription factor is indicated by circle on time axis. (B,C) Expression profiles of Mixer and Brachyury, and published targets selected for qPCR. Circled data points show onset times, dashed lines indicate qPCR sample points. (D,E) qPCR data relative to Odc1 for selected targets at 6.5 hpf and 9.5 hpf, comparing expression in transcription factor morpholino (MO)-injected versus control MO-injected embryos. Arrows show reduced expression in transcription factor knockdown. Data are two technical replicates from pairs of biological replicates: Mixer biological replicates are from a single clutch, Brachyury biological replicates are from different clutches. (F) Inhibition of in vitro translation of Mixer by Mixer MO showing the threshold concentration of morpholino able to block the translation of HA-tagged Mixer. Image published in: Collart C et al. (2014) © 2014. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |