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: Brain, behavior, and immunity

Article Title: Intermittent cytomegalovirus infection alters neurobiological metabolism and induces cognitive deficits in mice

doi: 10.1016/j.bbi.2023.12.033

Figure Lengend Snippet: Brain cell characterization flow panel.

Article Snippet: GBP2 , CF405M , Biorbyt , orb763167 , 1:200.

Techniques:

Journal: Heliyon

Article Title: Zhilong Huoxue Tongyu capsule inhibits rabbit model of hyperlipidemia and atherosclerosis through NF-κB/NLRP3 signaling pathway

doi: 10.1016/j.heliyon.2023.e20026

Figure Lengend Snippet: Details of antibodies.

Article Snippet: NFκB , Aviva Systems Biology, USA , arp38197_p050 , WB.

Techniques:

Journal:

Article Title: Pneumocystis Activates Human Alveolar Macrophage NF-?B Signaling through Mannose Receptors

doi: 10.1128/IAI.72.6.3147-3160.2004

Figure Lengend Snippet: AM NF-κB nuclear translocation in response to Pneumocystis organisms. (A) Time course for NF-κB nuclear translocation in AM from healthy individuals in response to Pneumocystis. The gel is from one healthy individual and is representative of three subjects examined. AM were incubated with unopsonized Pneumocystis for the indicated times, and EMSA was performed on AM nuclear extracts with a biotinylated (nonradioactive) NF-κB probe. Quantitative analysis (n = 3) performed by densitometry (values are means ± standard errors of the means) indicates optimal NF-κB nuclear translocation at 60 min. RU, relative units. (B) AM NF-κB nuclear translocation with different Pneumocystis MOIs. The gel is from one healthy individual and is representative of three subjects examined. AM were incubated with Pneumocystis (Pc) organisms for 1 h. Quantitative analysis performed by densitometry (values are means ± standard errors of the means) demonstrates optimal NF-κB nuclear translocation at an MOI of 5:1 (Pneumocystis/AM ratio). Unstim, unstimulated. (C) Detection of NF-κB subunits in response to Pneumocystis organisms (n = 5). EMSA with supershift from one healthy subject is shown and is representative of five subjects examined. The results demonstrate a supershift of NF-κB bands in the presence of antibodies directed against NF-κB p50 and p65 subunits but not p52, RelB, or c-Rel. (D) AM nuclear translocation of NF-κB detected by fluorescence microscopy with Cy3 (red) staining following a specific antibody recognizing the NF-κB p65 subunit. In unstimulated AM, the p65 staining is predominantly cytoplasmic, whereas incubation with Pneumocystis organisms for 1 h results in nuclear localization of NF-κB p65 staining. N, nucleus.

Article Snippet: The procedures for the supershift experiments were conducted as described above, with the exception that 2 μg of anti-p50, -p65, -p52, -RelB, and -c-Rel (Geneka, Montreal, Canada) antibodies or normal control serum was incubated with the nuclear cell extract for 30 min on ice before the addition of the biotin-end-labeled oligonucleotide probe.

Techniques: Translocation Assay, Incubation, Fluorescence, Microscopy, Staining