Journal: iScience

Article Title: Viral load dependent cellular heterogeneity in Trichomonas vaginalis at the single cell level

doi: 10.1016/j.isci.2025.114260

Figure Lengend Snippet: Trichomonasvirus species infection in T. vaginalis isolates The presence of TVV in the T. vaginalis isolates (ATCC 30236, ATCC 50143, ATCC 50148, ATCC PRA-98) was determined by mapping RNA-seq reads to Trichomonasvirus reference sequences obtained from the NCBI Virus Database using QIAGEN CLC Genomics Workbench (v20.0.3). (A) Summary of consensus coverage, average coverage, and total read counts. (B) Schematic representation of the consensus contig alignment for each Trichomonasvirus species. (C) Detection of Trichomonasvirus isolates using RT-PCR with virus species-specific primers, with PCR products separated on a 1% agarose gel.

Article Snippet: To investigate the distribution of Trichomonasvirus infection at single-cell resolution, scRNA-seq was performed on two samples derived from the same T. vaginalis isolate (ATCC 30236), obtained by splitting a single culture.

Techniques: Infection, RNA Sequencing, Virus, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis

Journal: iScience

Article Title: Viral load dependent cellular heterogeneity in Trichomonas vaginalis at the single cell level

doi: 10.1016/j.isci.2025.114260

Figure Lengend Snippet: Heterogeneous distribution of Trichomonasvirus in different T. vaginalis isolates (A) viral populations among the T. vaginalis isolates ATCC 50148. The plots show the presence of Trichomonasvirus species and their co-occurrence patterns within isolates. (B) Detection of viral dsRNA using the dsRNA-specific antibody 9D5 in different T. vaginalis isolates. Nuclei were stained with DAPI (blue), while viral dsRNA was detected using the 9D5 antibody conjugated with Alexa Fluor 594 (red). Images indicate the heterogeneity of viral dsRNA within the parasite. Scale bars = 10 μm.

Article Snippet: To investigate the distribution of Trichomonasvirus infection at single-cell resolution, scRNA-seq was performed on two samples derived from the same T. vaginalis isolate (ATCC 30236), obtained by splitting a single culture.

Techniques: Staining

Journal: iScience

Article Title: Viral load dependent cellular heterogeneity in Trichomonas vaginalis at the single cell level

doi: 10.1016/j.isci.2025.114260

Figure Lengend Snippet: Establishment of sub-lethal metronidazole exposure conditions for single-cell RNA sequencing in T. vaginalis Cell concentration curves of ATCC 30236 following exposure to metronidazole at 4, 6, and 8 μM, measured over a 24-h time course. All three concentrations led to complete growth arrest by ∼6 h post-treatment, though the degree of suppression varied slightly among them. We selected 6 μM (red line) as a representative sub-lethal condition, as it reflects an intermediate response and preserves >90% cell viability between 6 and 9 h. This concentration was therefore chosen to best capture the early transcriptional response window while maintaining cellular integrity. Data represent mean ± SEM from three independent biological replicates ( n = 3). Note: the y axis was truncated and compressed to allow both the high cell concentrations of the NC group and the suppressed growth of drug-treated groups to be visualized in the same panel.

Article Snippet: To investigate the distribution of Trichomonasvirus infection at single-cell resolution, scRNA-seq was performed on two samples derived from the same T. vaginalis isolate (ATCC 30236), obtained by splitting a single culture.

Techniques: RNA Sequencing, Concentration Assay

Journal: iScience

Article Title: Viral load dependent cellular heterogeneity in Trichomonas vaginalis at the single cell level

doi: 10.1016/j.isci.2025.114260

Figure Lengend Snippet: Viral load distribution and infection group composition in single-cell RNA-Seq Data This figure illustrates the distribution of viral RNA and the relative abundance of infection groups in two T. vaginalis single-cell RNA-seq samples (30236 and 30236_M). (A) Violin plots display normalized viral transcript levels (per 10 4 UMIs) across three infection states: High_infection, Infected, and Uninfected. (B) Stacked bar plots show the proportion of cells assigned to each infection group within each sample.

Article Snippet: To investigate the distribution of Trichomonasvirus infection at single-cell resolution, scRNA-seq was performed on two samples derived from the same T. vaginalis isolate (ATCC 30236), obtained by splitting a single culture.

Techniques: Infection, RNA Sequencing

Journal: iScience

Article Title: Viral load dependent cellular heterogeneity in Trichomonas vaginalis at the single cell level

doi: 10.1016/j.isci.2025.114260

Figure Lengend Snippet: Viral load-associated expression of functionally characterized gene groups in Trichomonas vaginalis Trend-based differential expression analysis was performed by stratifying virus-positive single cells into viral load quartiles (Q1-Q4) and comparing each bin to viral load-negative cells. (A) Drug susceptibility-associated genes, including flavodoxin-like fold proteins (TVAGG3_0053980, TVAGG3_0213430, TVAGG3_0585360, TVAGG3_0940660), thioredoxin-like proteins (TVAGG3_0038110, TVAGG3_0191900), disulfide oxidoreductase (TVAGG3_0154220), nitroreductase family proteins (TVAGG3_0195320, TVAGG3_0427810, TVAGG3_0695620), and pyruvate:ferredoxin oxidoreductase proprotein genes (TVAGG3_0282970, TVAGG3_0890230). (B) Ubiquitin-related genes (TVAGG3_0129220, TVAGG3_0221350, TVAGG3_0281680, TVAGG3_0586770, TVAGG3_0981600) are potentially involved in protein turnover and cellular stress responses. (C) Adhesion-associated genes, including TVAGG3_0540230, TVAGG3_0979910, TVAGG3_0163930, TVAGG3_0221780, and the heteropolysaccharide binding protein HPB2 (TVAGG3_0232100), were previously identified as adherence factors. Panels display per-bin expression distributions across viral load quartiles (Q1-Q4), overlaid with median trend lines.

Article Snippet: To investigate the distribution of Trichomonasvirus infection at single-cell resolution, scRNA-seq was performed on two samples derived from the same T. vaginalis isolate (ATCC 30236), obtained by splitting a single culture.

Techniques: Expressing, Quantitative Proteomics, Virus, Ubiquitin Proteomics, Binding Assay

Journal: iScience

Article Title: Viral load dependent cellular heterogeneity in Trichomonas vaginalis at the single cell level

doi: 10.1016/j.isci.2025.114260

Figure Lengend Snippet: Novel viral load-associated genes linked to stress response and chromatin regulation in T. vaginalis (A and B) Trend-based differential expression analysis revealed genes with positive correlations to viral load, including (A) myeloid leukemia factor (MLF)-related genes (TVAGG3_0389720, TVAGG3_0515090, TVAGG3_0703230, TVAGG3_0912280, TVAGG3_0969640) and (B) multiple histone-related genes (e.g., TVAGG3_0221660, TVAGG3_0309740, TVAGG3_0408090, TVAGG3_0473430). Boxplots show gene expression distributions across viral load quartiles (Q1-Q4), with median trend lines overlaid.

Article Snippet: To investigate the distribution of Trichomonasvirus infection at single-cell resolution, scRNA-seq was performed on two samples derived from the same T. vaginalis isolate (ATCC 30236), obtained by splitting a single culture.

Techniques: Quantitative Proteomics, Gene Expression