Fig. 5: Disruption of the ROCY1 locus dramatically alters the alkaline pH-stress-induced transcriptional response.
a Principal components (PC) 1 and 2 of cells infected with TgVEG WT, TgVEGΔROCY1, and TgVEGΔBFD1 expose to either control (●) or alkaline pH stress (▲) conditions for 48 h. N = 3 per strain and condition. b Number of transcripts with significant differences in abundance in high pH conditions compared to control conditions for TgVEG WT, TgVEGΔROCY1, and TgVEGΔBFD1 (|Log2 Fold Change| ≥ 1, Padj < 0.05). Of the 348 BIC-altered genes, the majority (261) were dependent upon both ROCY1 and BFD1 for their induction (based on a lack of significant induction (Padj>0.01) by pH in the knockout parasite line). Data from individual genes along with statistical analyses for pH responsiveness in all three strains is presented in Supplementary Dataset 2. c Heatmap of a subset of genes with significantly altered transcript abundance after exposure to high pH in T. gondii VEG WT parasites. All genes shown had BIC-altered transcript abundance by |log2FoldChange| > 2 and Padj<0.01. The complete set of BIC-regulated transcripts and their corresponding dependence on the presence of BFD1 or ROCY1 can be found in Supplementary Dataset 2 on the tab titled “348_BIC_INDUCED_GENES”. d, e Volcano plots of transcript abundance differences between wild type TgVEG and either TgVEGΔROCY1 (d) or TgVEGΔBFD1 (e) under BIC. The majority of transcripts that have significantly different abundance in each knockout line are those induced by BIC in wild type TgVEG. Red datapoints indicate transcripts with significant differences in abundance (Padj<0.01; |log2FC| > 1). This experiment was performed once. Data are provided as a Source Data File.
