Journal: mBio

Article Title: Impact of equilibrative nucleoside transporters on Toxoplasma gondii infection and differentiation

doi: 10.1128/mbio.02207-25

Figure Lengend Snippet: Transcriptomic response to acute TgENT1 depletion reveals a progressive, purine-centric stress program. TgENT1-mAID-HA tachyzoites (three biological replicates) were pre-treated for 48 h with vehicle or 500 µM IAA, then allowed to invade fresh Hs27 monolayers under the same conditions and harvested 24 h (day 1) or 48 h (day 2) after invasion. Poly(A) RNA was sequenced (Illumina NovaSeq; 150 bp PE), reads were mapped to the GT1 genome, and differential expression was analyzed with DESeq2 (|log₂FC| ≥ 1; false discovery rate [FDR] < 0.01). ( A ) Principal-component analysis (PCA). PC1 (40%) cleanly separates treated from control samples, whereas PC2 (28%) differentiates time points, indicating a robust, time-dependent transcriptional shift upon TgENT1 knockdown. ( B ) Heat map of the 50 most variable genes. Z -scored expression values highlight a concerted upregulation of purine- and nitrogen-metabolism genes (reds) and downregulation of cytoskeletal/motility genes (blues) that intensifies from day 1 to day 2. ( C ) Volcano plots of differentially expressed genes. Pink, upregulated; blue, downregulated; gray, non-significant. The magnitude and number of responsive genes expand markedly by day 2. The complete list of genes and fold change is presented in . ( D ) Overlap of DE genes between time points. Venn diagrams show 96 genes upregulated at both day 1 and day 2, and 58 genes consistently downregulated, underscoring a core TgENT1-dependent regulon that is sustained and amplified over time. ( E ) Functional enrichment analysis. Dot plot of the top gene ontology/KEGG terms (size = gene count; color = –log₁₀FDR). Upregulated sets are dominated by purine metabolism, nitrogen-compound metabolism, and cyano-/taurine-derivative pathways, whereas downregulated sets are enriched for microtubule-based movement and related processes, consistent with a shift from proliferation to metabolic stress response.

Article Snippet: RNA quality and concentration were confirmed, and poly(A)-selected mRNA libraries were subsequently prepared by Novogene.

Techniques: Quantitative Proteomics, Control, Knockdown, Expressing, Amplification, Functional Assay