Additional file 12: Figure S7. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

NvashA expression in animals with varied regiments of U0126 treatment. (A) NvashA expression in control animals, or in animals treated with U0126 continuously for 48 hours, or from 24 to 48 hpf. Unlike early stages when no NvashA expression could be detected (Fig. 3), NvashA expression was ultimately detected in U0126-treated animals by 48 hpf. Treatment with U0126 from 24 to 48 hpf reduced NvashA expression, but NvashA could be detected in many cells, albeit at reduced levels. (B) Levels of NvashA and Nvfgfa1 as detected by qPCR at late gastrula stage (48 hpf at 17 °C) in animals injected with the Nvfgfra MO or treated with U0126 or SU5402 from 24 to 48 hpf. Relative expression levels are compared to control MO- or DMSO-treated animals respectively. The red box defines 1.5 to −1.5 fold change region. Error bars are standard error. (TIF 11166 kb

Additional file 13: Figure S8. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

NvashA regulation of target genes in the embryonic ectoderm. Gene expression in NvashA morphants (A–C), control morpholino (D–F), and NvashA mRNA injected (G, H). Quantification below each image represents percent of embryos in each phenotypic class (see key in figure). All images except C and F are aboral views. C and F are oral views. Embryos were classified and quantified as the percent having normal expression, weak expression, or no expression.. The phenotypic class with the highest percentage of embryos is indicated. (TIF 21122 kb

Additional file 3: Figure S2. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

Changes in NvashA, Nvath-like, and NvsoxB2 expression following drug treatments or in with increased Notch activity. mRNA in situ hybridization for NvsoxB(2) (A) and NvashA (B) in DMSO-treated control animals grown to early planula stages (48 hpf at 22 °C). NvsoxB(2) (A’) and NvashA (B’) expression in same stage animal treated with U0126 from 24 to 48 hpf or treated with SU5402 from 24 to 48 hpf (B”). mRNA in situ expression of Nvath-like (C) and NvsoxB(2) (D) in control embryos injected with the venus mRNA. Expression of Nvath-like (C’) and NvsoxB(2) (D’) in animals injected with NvnotchICD:venus (the intracellular domain of the Notch receptor), which has been previously shown to hyperactivate Notch signaling. Embryos were classified and quantified as the percent having normal expression, weak expression, or no expression. The phenotypic class with the highest percentage of embryos is indicated. In C and D, the main figure panels are ectodermal focal planes, and insets show deeper focal planes used to confirm embryonic stage. All images are of lateral views with the oral side to the left. (TIF 34682 kb

Godetia amoena Lilja

原著和名: イロマツヨヒ科名: アカバナ科 = Onagraceae採集地: 千葉県 四街道市 千代田 (下総 四街道市 千代田)採集日: 1985/5/27採集者: 萩庭丈壽整理番号: JH026754国立科学博物館整理番号: TNS-VS-97675

Additional file 15: Table S7. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

Primer pairs used in this study for gene cloning. (XLS 73 kb

Additional file 10: Figure S5. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

Gene expression analyzed by quantitative polymerase chain reaction. Salt-and-pepper expressed genes as represented in Fig. 6. High-density gene expression profiles are represented by charts for all genes expressed at the blastula stage [24 hours post fertilization (hpf)] and/or gastrula stage (48 hpf) analyzed in this study. The y-axis indicates the relative fold change compared to unfertilized eggs. The x-axis indicates developmental time in hpf. Gene names as indicated in the top left corner and the Cp value in unfertilized eggs is indicated in the top right corner of each panel and was used to determine the presence of maternal transcripts in Fig. 6 (Cp > 34.00). Cp corresponds to the crossing point, also known as the cycle threshold (Ct) value. (PDF 599 kb

Additional file 9: Figure S4. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

Summary of animal hemisphere and aboral expression genes identified by U0126 array. mRNA in situ patterns are included in a manuscript currently in preparation. (PDF 39 kb

Additional file 1: Table S1. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

Dose-dependent effects of U0126 on Nvsprouty, Nvbrachyury, and Nv-fgfA1 expression. Dose-dependent effects of U0126 analyzed by in situ hybridization. Analyzed U0126 concentration as indicated in Row 1 and number of embryos with phenotype scored based on expansion/reduction of the domain of expression as indicated in the column on the right. (XLS 33 kb

Additional file 2: Figure S1. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

Summary of NvsoxB(2) RNA-seq data. Plot used data obtained from [33] from two duplicate RNA-seq data sets that generated transcriptomes over the first 19 hpf. Black trace shows average of two replicates. (JPG 471 kb

Additional file 4: Figure S3. of MAPK signaling is necessary for neurogenesis in Nematostella vectensis

NvAshA:Venus localization in U0126-treated animals. NvAshA:Venus protein was detected at high levels and with strong nuclear localization in U0126-treated animals. (TIF 16187 kb