Journal: PLOS Neglected Tropical Diseases

Article Title: A multiplex serological assay for quantification of arbovirus antibody kinetics and neutralizing antibodies

doi: 10.1371/journal.pntd.0013704

Figure Lengend Snippet: List of antigen names and suppliers used for multiplex bead-based assay development.

Article Snippet: Chikungunya non structural protein , CHIKV NSP , strain S27-African prototype , Cusabio , Togaviridae , CSB-EP810351CJAT.

Techniques: Multiplex Assay, Bead-based Assay, Virus, Control

Journal: PLOS Neglected Tropical Diseases

Article Title: A multiplex serological assay for quantification of arbovirus antibody kinetics and neutralizing antibodies

doi: 10.1371/journal.pntd.0013704

Figure Lengend Snippet: Normalized Median Fluorescence Intensity (nMFI) for IgG responses against antigens from chikungunya, Mayaro, O'nyong-nyong, and Ross River viruses in samples from France (green), Bangladesh (yellow), Senegal (pink), and the Philippines (purple). nMFI values are derived from median fluorescence intensity (MFI) using a Luminex INTELLIFLEX assay. The antigens assessed include virus-like particles (VLP), envelope glycoprotein E2 (E2), SNAP-tagged E2 (E2 + SNAP), non-structural protein (NSP) for chikungunya virus, and structural polyprotein (SP) for Ross River virus.

Article Snippet: Chikungunya non structural protein , CHIKV NSP , strain S27-African prototype , Cusabio , Togaviridae , CSB-EP810351CJAT.

Techniques: Fluorescence, Derivative Assay, Luminex, Virus

Journal: Journal of Virology

Article Title: Direct Activation of Endothelial Cells by SARS-CoV-2 Nucleocapsid Protein Is Blocked by Simvastatin

doi: 10.1128/JVI.01396-21

Figure Lengend Snippet: SARS-CoV2 nucleocapsid protein (NP) is a potent inducer of human endothelial cell activation. (a) HLMECs were incubated with SARS-CoV2 structural proteins (S, N, and E proteins; 1 μg/ml) and five nonstructural proteins (NSP1, NSP3, NSP5, NSP7, and NSP8; 1 μg/ml) for 8 h. (b) HLMECs were treated with 1 μg/ml of NP or 10 ng/ml of TNF-α for different incubation periods as indicated. (c) HLMECs were incubated with indicated concentrations of NP for 8 h. TNF-α at 10 ng/ml served as a positive control. (d) Different cultured cells, including mouse lung vascular endothelial cells (MECs), A549 cells, 293T cells, HUVECs, HAECs, HCAECs, HDMECs, and HLMECs, were treated with NP (1 μg/ml) for 8 h. The expression of ICAM-1, VCAM-1, and VE-cadherin was detected by Western blotting. β-Actin served as a loading control. (e) HLMECs were treated with PBS, NP (1 μg/ml), TNF-α (10 ng/ml), or lipopolysaccharide (LPS) (1 μg/ml) for 8 h. The total RNA was isolated and qPCR was performed for measuring the mRNA levels of TNF-α, ICAM-1, VCAM-1, MCP-1, and IL-6. (f) HLMECs were treated with PBS, NP (1 μg/ml), or TNF-α (10 ng/ml) for 8 h and cocultured with Zombie Red-labeled THP-1 cells for 1 h. After being washed, the adherent cells were imaged and quantitatively analyzed.

Article Snippet: SARS-CoV-2 NSP1 (97-095), NSP5 (10-116), NSP7 (97-096), and NSP8 (97-097) proteins were obtained from Prosci (Poway, CA).

Techniques: Activation Assay, Incubation, Positive Control, Cell Culture, Expressing, Western Blot, Control, Isolation, Labeling

Journal: Journal of cell science

Article Title: Derlin-1 promotes ubiquitylation and degradation of the epithelial Na + channel, ENaC.

doi: 10.1242/jcs.198242

Figure Lengend Snippet: Figure 7. HUWE1 is the ubiquitin E3 ligase that evokes ubiquitin-mediated

Article Snippet: The antibody of HUWE1 was purchased from Proteintech (Chicago, USA).

Techniques: Ubiquitin Proteomics

Journal: iScience

Article Title: A multimodal in situ framework for spatial characterization applied on SARS-CoV-2 live-attenuated vaccines in the nasal mucosa

doi: 10.1016/j.isci.2026.117730

Figure Lengend Snippet: Viral protein distribution and target cell tropism in the nasal mucosa at 2 dpi (A) SARS-NP positive areas in the olfactory epithelium were comparable between WT and OTS-206 inoculated hamsters and significantly larger than in OTS-228. (B) In the respiratory epithelium, larger SARS-NP positive areas were observed after OTS-206 than after WT inoculation. (C–E) Representative sections illustrate multifocal to diffuse SARS-NP labeling in epithelial cells of the olfactory epithelium (levels 2–3) and respiratory epithelium (levels 1–2; predominantly ciliated epithelial cells) following (C) WT and (D) OTS-206 inoculation. In contrast, (E) OTS-228 antigen was restricted to single cells or oligofocal clusters of few epithelial cells (overview, scale bars, 1 mm; detail, scale bars, 100 μm). (F) In the lamina propria of the olfactory mucosa, abundant viral antigen was detected after WT inoculation, whereas only minimal antigen was observed after OTS-206 and OTS-228. (G) WT led to prominent infection of numerous Bowman’s gland epithelial cells (black arrow) and only individual virus-positive glandular cells following OTS inoculation (scale bars, 100 μm). (H) The vomeronasal organ (black dotted contour) was affected in all WT-infected hamsters, but not in OTS-inoculated hamsters (scale bars, 100 μm). (I) SARS-NP antigen (black arrow) was detected in the cytoplasm of macrophages of the nasal-associated lymphoid tissue in all groups (scale bars, 100 μm). (J–L) Representative immunofluorescence images (63× magnification) of the olfactory epithelium identified (J) sustentacular cells (CK18, green) as the main target cell (SARS-NP, red) for all groups. (K) Few SARS-NP positive olfactory sensory neurons (Tuj1, cyan; magnified image, white dotted contour) were detected, while axon bundles in the lamina propria (overview image, white dotted contour) remained negative following OTS inoculation. (L) SARS-NP positive Bowman’s glands (CK18, green; white dotted contour). Kruskal-Wallis test followed by Dunn’s correction was performed (A, B, F). ∗ p < 0.05, ns = not significant. Mean with standard deviation is shown. SARS-NP: SARS-CoV nucleocapsid protein; CK18: cytokeratin peptide 18; Tuj1: neuron-specific class III beta-tubulin. n = 5 hamsters per group; 2 dpi; nasal levels 1, 2, and 3, with a total of 7 regions of interest, were analyzed.

Article Snippet: SARS-NP (1:3000, Rockland, 200–401-A50) IHC was performed as described above.

Techniques: Olfactory, Labeling, Infection, Virus, Tissue, Immunofluorescence, Standard Deviation

Journal: iScience

Article Title: A multimodal in situ framework for spatial characterization applied on SARS-CoV-2 live-attenuated vaccines in the nasal mucosa

doi: 10.1016/j.isci.2026.117730

Figure Lengend Snippet: Host gene responses in the olfactory epithelium at 2 (WT, OTS-206, OTS-228) and 5 dpi (OTS-228 only). RNA in situ hybridization of interferon beta 1 (IFN β1), retinoic acid inducible gene I (RIG I), myxovirus resistance protein 1 (Mx1), and tumor necrosis factor alpha (TNF α) combined with SARS-NP and Mx1 antigen detection (A) WT: SARS-NP (red) positive sustentacular cells (SCs) showed mainly moderate to abundant nuclear IFN-β1 RNA (purple), few to moderate RIG-I (green), and few Mx1 (yellow) RNA signals. Non-infected olfactory sensory neurons (OSNs) exhibited few to moderate cytoplasmic RIG-I and Mx1 RNA. (B) OTS-206: SARS-NP positive SCs showed moderate to abundant cytoplasmic IFN-β1 RNA. Virus antigen positive SCs and negative OSNs exhibited moderate RIG-I and Mx1 RNA. (C and D) OTS-228: At 2 dpi (C), RNA expression of all targets was below the detection limit. At 5 dpi (D), IFN-β1 remained undetectable, whereas few to moderate cytoplasmic RIG-I and Mx1 RNA were detected in both SARS-NP positive and negative SCs. (E–H) TNF-α RNA (cyan) was present at moderate to abundant levels at 2 dpi after WT and OTS-206 inoculation and restricted to SARS-NP positive SCs, showing (E) nuclear aggregation in WT and (F) predominantly cytoplasmic distribution in OTS-206. (G and H) OTS-228: TNF-α RNA was undetectable at 2 dpi (G), but at 5 dpi (H), few cytoplasmic signals were present in individual SARS-NP positive SCs. (I–L) Mx1 RNA and antigen. (I) WT antigen positive SCs showed absent or few Mx1 RNA (yellow) signals and minimal protein (blue), whereas non-infected SCs exhibited low to moderate levels of both. (J) OTS-206 antigen positive and- negative SCs with moderate to abundant Mx1 RNA and antigen. (K and L) OTS-228 antigen positive cells with minimal Mx1 protein at 2 dpi (K), and moderate to abundant Mx1 RNA and protein at 5 dpi (L). Dotted contours indicate SCs and OSNs in magnified images. SARS-NP: SARS-CoV nucleocapsid protein. n = 2 hamsters per group; 2 (all groups) and 5 dpi (OTS-228 only); 6 images (63× magnification) per animal taken from regions of interest were evaluated for location (nuclear, cytoplasmic, both) and abundance of RNA signals (few, moderate, abundant).

Article Snippet: SARS-NP (1:3000, Rockland, 200–401-A50) IHC was performed as described above.

Techniques: Olfactory, RNA In Situ Hybridization, Infection, Virus, RNA Expression

Journal: iScience

Article Title: A multimodal in situ framework for spatial characterization applied on SARS-CoV-2 live-attenuated vaccines in the nasal mucosa

doi: 10.1016/j.isci.2026.117730

Figure Lengend Snippet:

Article Snippet: SARS-NP (1:3000, Rockland, 200–401-A50) IHC was performed as described above.

Techniques: Plasmid Preparation, Virus, Indirect ELISA, Recombinant, RNAscope, Multiplex Assay, Software, Negative Control, Confocal, Laser-Scanning Microscopy

Journal: Structure (London, England : 1993)

Article Title: Structural insights into the assembly and regulation of 2'-O RNA methylation by SARS-CoV-2 nsp16/nsp10.

doi: 10.1016/j.str.2025.03.009

Figure Lengend Snippet: Figure 3. Putative ligand binding pocket of nsp16 (A) Snapshots of various ligands that bind to the pocket in different nsp16/nsp10 structures: adenosine (PDB: 6WKS),8 m7GDP (PDB: 6WQ3),9

Article Snippet: The following antibodies were used: SARS-CoV-2 nsp10 (ProSci, 9179), SARS-CoV-2 nsp16 (ProSci, 9271), anti-rabbit IgG HRP secondary (Cell Signaling, 7074).

Techniques: Ligand Binding Assay

Journal: Structure (London, England : 1993)

Article Title: Structural insights into the assembly and regulation of 2'-O RNA methylation by SARS-CoV-2 nsp16/nsp10.

doi: 10.1016/j.str.2025.03.009

Figure Lengend Snippet: Figure 5. Sampling oligomeric conforma- tions in solution (A) Coomassie staining of crosslinked nsp16/ nsp10 (±RNA) on SDS-PAGE, with protein bands labeled as 1–6. (B) Immunoblotting with nsp16 antibody shows nsp16 in bands 2, 4–6, and part of wider band 3 (3A). (C) Immunoblotting with nsp10 antibody detects nsp10 in bands 1, 5, 6, and parts of wider band 3 (3B and 3C). Cartoon representations of their olig- omeric states are shown below (blue circle: nsp10, green square: nsp16). (D) Coomassie staining of crosslinked quintuple mutant (±RNA), protein bands labeled as 1–6. (E and F) Immunoblotting with nsp16 and nsp10 antibodies, respectively.

Article Snippet: The following antibodies were used: SARS-CoV-2 nsp10 (ProSci, 9179), SARS-CoV-2 nsp16 (ProSci, 9271), anti-rabbit IgG HRP secondary (Cell Signaling, 7074).

Techniques: Sampling, Staining, SDS Page, Labeling, Western Blot, Mutagenesis

Journal: Structure (London, England : 1993)

Article Title: Structural insights into the assembly and regulation of 2'-O RNA methylation by SARS-CoV-2 nsp16/nsp10.

doi: 10.1016/j.str.2025.03.009

Figure Lengend Snippet: Figure 6. Oligomerization via nsp16-nsp16 interfaces and MP analysis (A) nsp16-nsp16 interface 1 in the current struc- ture. (B) Previously reported interface 2 (PDB: 7JYY). (C) An overlay revealing distinct interfaces on opposite faces of the nsp16/nsp10 heterodimer. (D and E) Mass photometry of the (D) WT apo (gray) and RNA-bound (1:1 M ratio, dark green) and (E) quintuple mutant.

Article Snippet: The following antibodies were used: SARS-CoV-2 nsp10 (ProSci, 9179), SARS-CoV-2 nsp16 (ProSci, 9271), anti-rabbit IgG HRP secondary (Cell Signaling, 7074).

Techniques: Mutagenesis