Gene expression of axenically-isolated clinical Entamoeba histolytica strains and its impact on disease severity of amebiasis
Fig 5
Transcriptome profiling of the E. histolytica laboratory strain (HM-1:IMSS clone 6).
(A) Experimental work flow for the three different culture conditions: HM-1 (in vitro), maintained under in vitro culture conditions for many years; HM-1 (virulent), maintained under virulent conditions by routinely passaging through an animal liver every 3 months; HM-1 (liver), collected just after liver challenge with strain HM-1. (B) The proportion of liver abscess weight to whole liver weight for strain HM-1 (virulent). The proportion was positively correlated with the dose of HM-1 (virulent), but the linear relationship of HM-1 (virulent) was relatively weak compared with that of the highly virulent Ax19 strain. (C) 2D plot of principle component analysis (PCA) of RNA-seq reads from the three different culture conditions for strain HM-1, using the two different culture conditions for the highly virulent Ax19 strain as reference gene profiles. Each data point represents a read, analyzed in triplicate. The expression profile of HM-1 (in vitro) was clearly different from that of HM-1 (virulent). The difference was greater than the changes induced by liver challenge (HM-1 (virulent) vs HM-1 (liver)). Moreover, the expression profile of HM-1 (virulent) was also distinct from that of ALA19. (D) Heat map of the three different culture conditions for the HM-1 strain. The expression profile of HM-1 (in vitro) was clearly different from those of HM-1 (virulent) and HM-1 (liver). (E and F) Volcano plots showing each differentially expressed gene (DEG) for the E. histolytica laboratory strain. Although there were 6,309 DEGs identified between HM-1 (in vitro) and HM-1 (virulent), there were no more than 565 DEGs identified between HM-1 (virulent) and HM-1 (liver). (G) Venn diagram of the 85 Ax19 environment-specific DEGs and the 565 HM-1 environment-specific DEGs. The number of overlapping genes was 21. (H) GO function analysis of the 565 HM-1 environment-specific DEGs.