Journal: bioRxiv

Article Title: A Suite of Eight Toxoplasma gondii Effectors Cooperates to Activate the Non-canonical NF-κB Pathway

doi: 10.64898/2026.03.12.711255

Figure Lengend Snippet: A) Top-scoring transcription factors predicted to regulate the MYR1-dependent upregulated genes in HFF cells relative to uninfected controls were identified by analysis of previously published data using Enrichr. B-C) Representative images (top) and quantitative analysis (bottom) for RelB (B) and p52 (C) nuclear accumulation in HFFs infected with RH (WT), Δ myr1 or Δ myr1 ::MYR1 complement parasites. Twenty-four hours post-infection, cells were fixed and labeled with DAPI (blue), anti-GAP45 (red), and either anti-RelB or anti-p52 (green). Scale bars = 10 µm. Plots display the nuclear-to-cytoplasmic signal ratios for at least 300 infected cells (red arrows) per condition; uninfected cells are indicated by white arrows. Data from three independent experiments were combined for analysis. The horizontal dashed lines indicate the mean. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparison test; ****P < 0.0001, ns = not significant.

Article Snippet: The following primary antibodies were used: rabbit anti-RelB (1:300; Proteintech, 25027-1-AP), mouse anti-NFκB p52 (1:100; Santa Cruz Biotechnology, sc-7386), and mouse anti-GAP45 or rabbit anti-GAP45 (kindly provided by Dr D. Etheridge, University of Georgia, USA).

Techniques: Infection, Labeling, Comparison

Journal: bioRxiv

Article Title: A Suite of Eight Toxoplasma gondii Effectors Cooperates to Activate the Non-canonical NF-κB Pathway

doi: 10.64898/2026.03.12.711255

Figure Lengend Snippet: A-B) Representative images (top) and quantitative analysis (bottom) for RelB (A) and p52 (B) nuclear accumulation in MEFs infected with RH (WT), Δ myr1 or Δ myr1 ::MYR1 complement parasites. Twenty-four hours post-infection, cells were fixed and labeled with DAPI (blue), anti-GAP45 (red), and either anti-RelB or anti-p52 (green). Scale bars = 10 µm. Plots display the nuclear-to-cytoplasmic signal ratios for at least 300 infected cells (red arrows) per condition; uninfected cells are indicated by white arrows. Data from three independent experiments were combined for analysis. The horizontal dashed lines indicate the mean. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparison test; ****P < 0.0001, ns = not significant.

Article Snippet: The following primary antibodies were used: rabbit anti-RelB (1:300; Proteintech, 25027-1-AP), mouse anti-NFκB p52 (1:100; Santa Cruz Biotechnology, sc-7386), and mouse anti-GAP45 or rabbit anti-GAP45 (kindly provided by Dr D. Etheridge, University of Georgia, USA).

Techniques: Infection, Labeling, Comparison

Journal: bioRxiv

Article Title: A Suite of Eight Toxoplasma gondii Effectors Cooperates to Activate the Non-canonical NF-κB Pathway

doi: 10.64898/2026.03.12.711255

Figure Lengend Snippet: T. gondii activates the non-canonical NF-κB pathway through MYR1-dependent TRAF3 depletion and NIK stabilization. (A) Representative images (top) and quantitative analysis (bottom) for RelB nuclear accumulation in HFFs infected with RH (WT) parasites over a 24-h time course. At 6, 12, 18, and 24 h post-infection, cells were fixed and labeled with DAPI (blue), anti-GAP45 (red), and anti-RelB (green). Scale bars = 10 µm. Plots display the nuclear-to-cytoplasmic intensity ratios for at least 300 infected cells (red arrows) per condition; uninfected cells are indicated by white arrows. Data from three independent experiments were combined for analysis. The horizontal dashed lines indicate the mean. Statistical significance was determined using a one-way ANOVA with Tukey’s multiple comparison test; ***P < 0.001, ****P < 0.0001, ns = not significant. (B) Immunoblot analysis (top) and corresponding quantification (bottom) of TRAF3, NIK, p100 phosphorylation, and p100-to-p52 processing in HFFs that were uninfected (UI) or infected with RH (WT), Δmyr1 or Δmyr1 ::MYR1 complement parasites. Lysates collected 24 h post-infection were resolved by SDS-PAGE and probed with specific primary antibodies; β-tubulin served as a loading control. Band intensities were measured using ImageJ and normalized to β-tubulin. Data are presented as mean ±SD from three biological replicates. Statistical significance for all panels was determined using a one-way ANOVA with Tukey’s multiple comparison test; **P < 0.01, ***P < 0.001, ****P < 0.0001, ns = not significant

Article Snippet: The following primary antibodies were used: rabbit anti-RelB (1:300; Proteintech, 25027-1-AP), mouse anti-NFκB p52 (1:100; Santa Cruz Biotechnology, sc-7386), and mouse anti-GAP45 or rabbit anti-GAP45 (kindly provided by Dr D. Etheridge, University of Georgia, USA).

Techniques: Infection, Labeling, Comparison, Western Blot, Phospho-proteomics, SDS Page, Control

Journal: Nature Communications

Article Title: G-quadruplex as an essential structural element in cytomegalovirus replication origin

doi: 10.1038/s41467-024-51797-6

Figure Lengend Snippet: a Scheme for the pull-down assay to identify viral and cellular proteins binding to ER-I G4 (see Methods). b , c HF cells were infected with HCMV at an MOI of 1. Cell lysates were prepared at 120 h and used for pull-down assays. Biotinylated ODNs (0.5 μg), pre-incubated to form G4 or i-motif structures (see the Methods), were incubated with 50 μg of cell lysates and immobilized on streptavidin beads. After washing, the proteins were eluted and analyzed by immunoblotting with specific antibodies for hnRNP K, IE1/IE2, p52 (encoded by UL44), and UL84. Input (1/10), 5 μg of cell lysates. (-), no biotinylated ODNs added ( b ). The relative binding efficiency of proteins [input (1/10)%] to different ODNs in three independent experiments is also indicated in ( c ). The data are shown as the mean ± SD. Source data are provided as a file.

Article Snippet: Anti-UL44 (p52) and anti-UL84 mouse MAbs were purchased from Virusys.

Techniques: Pull Down Assay, Binding Assay, Infection, Incubation, Western Blot