Supplementary MaterialsFigure 1source data 1: Quantifications and statistical analysis of larval cells related to Figure 1A. Amiodarone hydrochloride SEM are reported in the source data file and are highlighted in yellow, magenta, blue, green, and cyan, respectively.DOI: http://dx.doi.org/10.7554/eLife.20145.007 elife-20145-fig2-data1.xlsx (39K) DOI:?10.7554/eLife.20145.007 Figure 3source data 1: Quantifications and statistical analysis of larval cells corresponding to Figures 3A. Exact values of N, mean, median, SD, and SEM are reported in the source data file and are highlighted in yellow, magenta, blue, green, and cyan, respectively.DOI: http://dx.doi.org/10.7554/eLife.20145.009 elife-20145-fig3-data1.xlsx (39K) DOI:?10.7554/eLife.20145.009 Figure 3source data 2: Quantifications and statistical analysis of larval cells corresponding to Figure 3B. DOI: http://dx.doi.org/10.7554/eLife.20145.010 elife-20145-fig3-data2.xlsx (38K) DOI:?10.7554/eLife.20145.010 Figure 3source data 3: Quantifications and statistical analysis of larval cells corresponding to Figure 3C. DOI: http://dx.doi.org/10.7554/eLife.20145.011 elife-20145-fig3-data3.xlsx (33K) DOI:?10.7554/eLife.20145.011 Figure 4source data 1: Quantifications and statistical analysis of larval cells corresponding to Figure 4B. Exact values of N, mean, median, SD, and SEM are reported in the source data file and are highlighted in yellow, magenta, blue, green, and cyan, respectively.DOI: http://dx.doi.org/10.7554/eLife.20145.014 elife-20145-fig4-data1.xlsx (39K) DOI:?10.7554/eLife.20145.014 Figure Amiodarone hydrochloride 5source data 1: Quantifications and statistical analysis of proliferation of larval cells corresponding to Figure 5B. Exact values of N, mean, median, SD, and SEM Amiodarone hydrochloride are reported in the source data file and are highlighted in yellow, magenta, blue, green, and cyan, respectively.DOI: http://dx.doi.org/10.7554/eLife.20145.017 elife-20145-fig5-data1.xlsx (46K) DOI:?10.7554/eLife.20145.017 Figure 6source data 1: Quantifications and statistical analysis of larval cells corresponding to Figure 6D. Exact values of N, mean, median, SD, and SEM are reported in the source data file and are highlighted in yellow, magenta, blue, green, and cyan, respectively.DOI: http://dx.doi.org/10.7554/eLife.20145.020 elife-20145-fig6-data1.xlsx (39K) DOI:?10.7554/eLife.20145.020 Supplementary file 1: Mass spectrometry: CFS inventory. Relative abundance of protein species (spectral count) within the 60C80% ammonium sulfate fraction of the gene are present in the genomes of a subset of human intestinal bacteria, and we show that two of the corresponding proteins share BefAs capacity to induce cell expansion in zebrafish. Results The microbiota is required for normal expansion of the larval cell mass To investigate a possible role for the microbiota in pancreas development and specifically in cell expansion, we quantified total ?cells in GF and conventionally reared (CV) fish (diIorio et al., 2002) at 3, 4, 5 and 6 dpf (Figure 1A, Figure 1source data 1). The number of cells in CV fish increased steadily from 3 to 6 dpf (Figure 1A). However, the average number of cells in GF fish remained static over this time (Figure 1A). Furthermore, at 6 dpf, the overall structure of cells within the primary islet also appeared much less densely packed in GF than in CV fish (Figure 1B). This effect is not likely to be due to changes in initial differentiation of the cell population since the total number of cells is not different between GF and CV fish at 3 dpf (Figure 1A), a time at which exposure to bacteria is also limited. Open in a separate window Figure 1. The microbiota are necessary for regular expansion from the larval cell mass.(A) Final number Rabbit Polyclonal to ATG4A of cells per larva in GF (white box plots) and CV (gray box plots) seafood at 3, 4, 5 and 6?dpf.?With this, and in every subsequent numbers, CV data are demonstrated in grey package plots, and GF data, or similar treatment groups statistically, are demonstrated in white package plots. In every relevant sections and remaining numbers, box storyline whiskers represent the 95% self-confidence interval of the info set. Single element ANOVA indicates that gnotobiology of the fish was significant in determining the number of cells present (F7=9.01, p=1.45e?8). Labels a, ab and b indicate the?results of post hoc means testing (Tukey). The difference between GF and CV cell counts became significant at 6 dpf (t=?5.91, p 0.001). (B) Representative 2D slices from confocal scans through the primary islets of 6 dpf CV and GF larvae. Each slice is taken from the approximate center of the islet structure. Insulin promoter expressing cells are in green and nuclei are blue. Scale bar = 40?M. (C) The?average amount of glucose (pmol) per larva aged 6 dpf (* t17=?3.65, p 0.01). DOI: http://dx.doi.org/10.7554/eLife.20145.003 Figure 1source data 1.Quantifications and statistical analysis of larval cells corresponding to Figure 1A. Exact values of N, mean, median, SD, and SEM are.