Fig. 2

Transcriptomic analysis on vegetative, streaming, mound, and fruiting body stages of D. discoideum reveals unicellular and multicellular specific gene expression patterns. a A heatmap of gene expression of all 13,267 genes at different stages of D. discoideum reveals gene sets that increase, decrease, and remain constant throughout progression to multicellular stages. Scale bar is displayed below. a and b indicate independent replicates. b Principle components analysis (PCA) of gene expression demonstrates that biological replicates cluster together and unicellular, vegetative stage clusters separately from the three multicellular stages. c Representative stage-specific gene expression tracks of known stage-specific genes. d Real-time qPCR validation of representative multicellular and unicellular gene expression from RNAseq datasets. This graph represents the mean ± the standard deviation of three biological replicates performed in triplicate. e Stage by stage Gene Ontology (GO) comparison of differentially regulated genes reveals that classes of genes which correlate with multicellularity are misregulated when comparing Vegetative (V) to Streaming (S) or Fruiting body (F) stages. f A Bland-Altman plot of D. discoideum RNAseq datasets reveals genes which are upregulated and downregulated between unicellular and both multicellular stages. Red dots represent significantly differentially expressed genes, black dots represent not significantly differentially expressed genes. g The top ten GO terms of unicellular enriched (top) or multicellular enriched (bottom) processes reveals genes involved in essential processes are enriched in unicellular D. discoideum and processes required for development, differentiation, communication, and adhesion are enriched in multicellular D. discoideum. Heat maps of transcription factors (h), and chromatin modifying enzymes (i), that display unicellular or multicellular enriched expression are displayed here. An analysis of all transcription factors and all chromatin-modifying enzymes identified in D. discoideum is presented in Additional file 1: Figure S10