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Figure S5. Temperature and MO Dosage Effects on Brachyury Phenotype and Immune Response Related Gene Transcription, Related to Figure 7 (A) WMISH for tp53 of late tailbud embryos injected with 4.5 or 18 ng of the t/t2 MO mix and developed at 22 ºC or 28.5ºC. *, Remark: Increasing incubation temperature also slightly up-regulated tp53 in the absence of any MO (confirmed by RT-qPCR, data not shown). (B) Multi-probe WMISH for various mesoderm cell lineage and derivative markers (cav1, notochord; hoxd8, pronephros; myh6, heart; tal1, ventral blood island; tbx6, paraxial mesoderm) and single WMISH for tp53, tp53inp1 and c3ar1 of mid-tailbud (stage 26) and late tailbud embryos (stage 34) injected with 6 or 18 ng of the t/t2 MO mix. White arrowheads point to the expression domains of tbx6 and cav1 that were not maintained in embryos without functional Brachyury. (C) WMISH for tp53inp1 on wild-type (or t/t2 heterozygous) and t/t2 null mutant embryos as well as embryos injected with 1 and 3 ng MO. The embryos were developed to late tailbud stage 34 at 25-26ºC. Scale bar, 0.5 mm. Image published in: Gentsch GE et al. (2018) © 2018 The Francis Crick Institute. 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 Gene Synonyms Species Stage(s) Tissue tp53.L p53, Trp248, tumor suppressor p53, Xp53, Xrel3 X. laevis Throughout NF stage 33 and 34 head pharyngeal region tp53inp1.L X. laevis Throughout NF stage 33 and 34 head pharyngeal region hoxd8.L hox5.4, hoxd-8 X. laevis Sometime during NF stage 26 to NF stage 33 and 34 pronephric tubule cav1.L bscl3, Cav-1, cav1-a, cav1-b, caveolin-1, cgl3, vip21 X. laevis Sometime during NF stage 26 to NF stage 33 and 34 notochord tbx6.L XTbx6 X. laevis Sometime during NF stage 26 to NF stage 33 and 34 tail bud myh6.L aMHC, ck231, MHC, MHC-alpha, MHCa, MHCalpha, muz, muzak, XCMHC X. laevis Sometime during NF stage 26 to NF stage 33 and 34 heart tal1.L bHLHa17, stem cell leukaemia, tal-1, tcl5, xscl X. laevis Sometime during NF stage 26 to NF stage 33 and 34 ventral blood island Image source: Published Larger Image Printer Friendly View

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