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Fig. 4 | Genome Biology

Fig. 4

From: Role of epigenetics in unicellular to multicellular transition in Dictyostelium

Fig. 4

Genes identified as accessible and expressed in multicellular but not unicellular stages and organisms are necessary for multicellularity in D. discoideum. a Representative images illustrating that knock-out of the multicellular genes, hdaD, srfA, ammr1, and smcl1, but not the unicellular gene, hspH2, results in multicellularity delays. Representative images are shown at 8, 16, 21, 25, and 41 h, and the stage is noted in the bottom right hand corner of each image. Scale bars are 20 microns. b Knockout of multicellular genes delays multicellularity (upper graph) and total transition to fruiting body stage (lower graph) but knockout of a unicellular gene does not. This graph represents the mean ± the standard error of the mean of three independent biological replicates performed in triplicate. Ns: not significant, *: p < 0.05, **: p < 0.005, ***: p < 0.001, ****: p < 0.0001 as assessed by multiple comparison one-way ANOVA analysis. c Knockout of srfA but not hdaD or ammr1 caused a reduction in the diameter of the fruiting body as a proxy for the number of cells in each multicellular organism. Knockout of hspH2 caused an increase in the diameter of the fruiting body. This graph represents the mean ± the standard error of the mean of three independent biological replicates performed triplicates. Ns: not significant, *; p < 0.05, **; p < 0.005 as assessed by multiple comparison one-way ANOVA analysis. d Knock-out of multicellular genes: hdaD, ammr1, and smcl1 had no effect on chemotaxis while knock-out of srfA and unicellular gene hspH2 decrease the chemotaxis capacity of D. discoideum. The bar graph represents the mean ± the standard error of the mean of three biological replicates performed in triplicate. The number of cells which migrated towards the 30° segment containing the location where 250 μM folate was placed was counted. Ns: not significant, *: p < 0.05, **: p < 0.005 as assessed by multiple comparison one-way ANOVA analysis. e Knock-out of unicellular and multicellular genes had no effect on cell viability as assessed by PI staining. All knock-out strains responded similarly to the WT strain in response to 95 °C heat shock for 50 seconds. Each bar represents the mean ± the standard deviation of three biological replicates. Ns: not significant as assessed by one-way ANOVA analysis

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