Gene expression data

Results obtained from the NanoString analysis of gene expression during zebrafish development (raw data, normalised data, and analysis)

Codeset information

CodeSet information (gene names, accession numbers, target sequences) for the NanoString probe set used in this study

Additional file 7: of Single swim sessions in C. elegans induce key features of mammalian exercise

Primers used for qPCR. (DOCX 16 kb

Additional file 1: of Single swim sessions in C. elegans induce key features of mammalian exercise

Changes in specific oxidative stress response transcripts accompany swim exercise in C. elegans. (A–L) qPCR results in N2 animals before and at different time points post-exercise for oxidative stress reporter genes (n = 5 independent trials). Note the different y-axis scales between figure panels (particularly K and L). hsp-16.2 and hsp-16.41 are documented to be induced under both oxidative stress and heat shock. We calculated relative expression by normalization to reference genes followed by normalization to the time point before exercise. We used paired two-tailed Student’s t tests to compare relative expression of control versus exercise samples at each time point. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. (PDF 260 kb

Effects of Dll4 in hematopoiesis.

<p>(A) Flow cytometry analysis of CD41 expression in WT, Dll4^+/− and Dll4^−/− EBs from day 3.5 to day 6 of differentiation. (B) Number of CFU-EryP; (C) CFU-definitive erythroid (EryD) and (D) total definitive CFUs (include CFU-EryD, -macrophage, -granulocyte, -granulocyte//macrophage and -erythroid/granulocyte/macrophage) generated by WT, Dll4^+/− and Dll4^−/− EB cells at days 4, 5, 6 and 8 of differentiation. (E–F) Number of CFU-EryP (E) and total definitive CFUs (F) generated by YS cells from WT, Dll4^+/− and Dll4^−/− mouse embryos at E8.0, E8.5 and E9.0. Each symbol represents individual YS and horizontal lines indicate the mean value. (G) Number of CFU-EryP generated by WT and Dll4^+/− cells at day 5 of differentiation on parental OP9 cells (OP9-WT) or OP9 cells transduced either with retroviral vectors containing Dll4 (OP9-Dll4), Dll1 (OP9-Dll1) or only IRES-eGFP sequences (OP9-Empty). ND not determined. *<i>P</i><.05, **<i>P</i><.01, ***<i>P</i><.001.</p

Analysis of Flk1 expression and BL-CFC potential of WT and Dll4 mutant EBs.

<p>(A) Flow cytometry analysis of Flk1 expression in WT, Dll4^+/− and Dll4^−/− EBs from day 2 to day 6 of differentiation. (B) Number of blast colonies generated by WT, Dll4^+/− and Dll4^−/− EB cells from day 2 to day 5 of differentiation. (C) Number of blast colonies generated by sorted Flk1 cells derived from day 2.5 WT and Dll4^+/− EBs and from day 3.5 Dll4^−/− EBs. (D) Number of blast colonies generated by day 3 EBs (upper) or sorted Flk1 cells (bottom) differentiated from WT and Dll4^−/− ES cells transduced with empty (pMigR1) or recombinant virus (pMigR1-Dll4). (E) Representative flow cytometry dot-plot of Flk1/PdgfR-α expression from day 3 to day 4 of WT, Dll4^+/− and Dll4^−/− EBs (Flk1⁺/PdgfR-α⁺: day 3 - WT and Dll4^+/−<i>versus</i> #7, 0.00060<<i>P</i><0.0406; Flk1⁺/PdgfR-α⁻: day 3 to day 4 - WT and Dll4^+/−<i>versus</i> #7, 0.0002<<i>P</i>< 0.0094; day 4 - WT <i>versus</i> Dll4^+/−, <i>P</i> = 0.0457). ND not determined. *<i>P</i><.05, **<i>P</i><.01, ***<i>P</i><.001.</p

Generation and characterization of Dll4 mutant EBs.

<p>(A) Semi-quantitative RT-PCR analysis of <i>Dll4</i> expression in EBs generated from WT-R1 ES cells, from day 0 to day 8. Data representative of three independent experiments. (B) PCR genotyping of WT and mutant Dll4 alleles of WT, Dll4^+/− and three Dll4^−/− (#7, #9i and #10i) ES cell lines. (C) Semi-quantitative RT-PCR analysis of <i>Dll4</i> expression in WT, Dll4^+/− and Dll4^−/− EBs with 3 days of differentiation. (D) Representative flow cytometry dot-plot of SSEA-1/Flk1 expression in WT, Dll4^+/− and Dll4^−/− EBs from day 3 to day 4.5. (E) Immunofluorescence detection of E-cadherin (membrane bright staining), counterstained with DAPI (nuclear gray staining), of WT, Dll4^+/− and Dll4^−/− (#7) EBs with 3 days of differentiation. (F) Semi-quantitative RT-PCR analysis of <i>Rex1</i>, and genes involved in the formation of germ layers in WT, Dll4^+/− and Dll4^−/− EBs from day 0 to day 8 of differentiation. Data representative of three independent experiments. Scale bar represents 10 µm.</p

Differential effects of Dll4 on the emergence of SMCs and cardiomyocytes.

<p>(A–C) Representative immunofluorescence images for αSMA (cytoplasmic positive fibers), counterstained with DAPI (nuclear staining), of day 3 EB-derived cells cultured for 5 days in monolayer culture. (D) Percentage of αSMA⁺ cells obtained from day 3 EB-derived cells of WT, Dll4^+/− and Dll4^−/− cell lines. (E) Percentage of beating EBs in suspension culture from day 5 to day 9 of differentiation. (F) Semi-quantitative RT-PCR analysis of <i>Nkx2-5</i> in WT, Dll4^+/− and Dll4^−/− EBs from day 3 to day 8 of differentiation. Scale bar represents 10 µm. *<i>P</i><.05, **<i>P</i><.01, ***<i>P</i><.001.</p