Journal: The FASEB Journal

Article Title: ChREBP ‐β Exacerbates Renal Tubular Disorders Caused by Fructose via ATF4

doi: 10.1096/fj.202600490R

Figure Lengend Snippet: Effects of ChREBP‐β renal tubular‐specific overexpression on the kidney of mice. (A) Schematic demonstration. (B) Representative demonstration for PCR‐based genotyping. (C) Real time PCR analysis for ChREBP‐β overexpression efficiency in the kidney. (D,E) GTT of mice and the area under the curve statistics. (F) Representative images of HE and Tunel‐stained kidney sections. Scale bars = 100 μm. (G) Representative TEM images showing the brush border area in the kidneys of mice. Scale bars = 1 μm. n = 6; all data are expressed as mean ± SD. * p < 0.05.

Article Snippet: DNA extraction was performed using the One Step Mouse Genotyping Kit (PD101‐01, Vazyme) according to the manufacturer's instructions.

Techniques: Over Expression, Real-time Polymerase Chain Reaction, TUNEL Assay, Staining

Journal: NPJ Vaccines

Article Title: A system to rapidly develop a bunyavirus pseudotyped virus neutralisation assay for pandemic preparedness

doi: 10.1038/s41541-025-01278-8

Figure Lengend Snippet: RVFV ( A ) and CCHFV ( B ) M segment in open reading frame expression, with arrow indicating the four potential RVFV GnGc start codons. MLD mucin-like domain. Recombinant VSV or HIV-1-based vectors expressing the firefly luciferase gene were used to generate PV for RVFV ( C ) or CCHFV ( D ). The PV was titrated on Vero cells (VSV-based) or CRFK cells (HIV-1-based) and titres expressed as 50% tissue culture infectious dose (TCID 50 ) per mL determined by the detection of luminescence and using Spearman–Karber formula. The graph shows the mean value of three independent experiments ± standard error of the mean.

Article Snippet: Cells were then incubated with 50 μL/well of the primary antibody anti-RVFV nucleoprotein mouse monoclonal antibody (M977, The Native Antigen Company) diluted 1:1000 in blocking buffer, for 1 h at room temperature.

Techniques: Expressing, Recombinant, Luciferase, Endpoint Dilution Assay

Journal: NPJ Vaccines

Article Title: A system to rapidly develop a bunyavirus pseudotyped virus neutralisation assay for pandemic preparedness

doi: 10.1038/s41541-025-01278-8

Figure Lengend Snippet: Increasing amounts of expression plasmid for the native sequence of RVFV GnGc glycoproteins ( A ) or human-codon optimised sequences ( B ) were used to transfect HEK293T/17 cells using either PEI or FugeneHD to produce VSV-based PV. Comparison of the original (wt) against codon optimised (CO) CCHFV GnGc sequence for the production of VSV-based PV using PEI transfection reagent and increasing amount of expression plasmid ( C ). CCHFV-PV titres using the codon-optimised GnGc sequence were compared between the two transfection reagents ( D ). Titres are reported as 50% tissue culture infectious dose (TCID 50 ) per mL. Results for three or four independent productions are reported as mean titre ± standard error of the mean. The coefficient of variation (CV%) is reported below each graph. Immunoblot of the VSV-based pseudotyped particles using anti-RVFV Gn ( E ) or anti-CCHFV Gn ( F ) monoclonal antibodies. The predicted size of the monomer is indicated in the blot.

Article Snippet: Cells were then incubated with 50 μL/well of the primary antibody anti-RVFV nucleoprotein mouse monoclonal antibody (M977, The Native Antigen Company) diluted 1:1000 in blocking buffer, for 1 h at room temperature.

Techniques: Expressing, Plasmid Preparation, Sequencing, Comparison, Transfection, Western Blot, Bioprocessing

Journal: NPJ Vaccines

Article Title: A system to rapidly develop a bunyavirus pseudotyped virus neutralisation assay for pandemic preparedness

doi: 10.1038/s41541-025-01278-8

Figure Lengend Snippet: Schematic of the timeline for the production of functional PVs once the viral sequence is published and assessment of usable PV. Created in BioRender https://BioRender.com/9cj39l1 ( A ). Human-codon optimised sequences of Rift Valley fever virus, (RVFV), Dabie bandavirus (BADV), Oropouche virus (OROV) and Crimean-Congo haemorrhagic fever virus (CCHFV) GnGc sequences were used to produce PV and titrated on Vero (black) or Huh-7 (red) cells ( B ). PV titres are reported based on luciferase activity as TCID 50 /mL. Each point indicates an independent production of the PVs. The significance between the titration on Vero versus Huh7 was determined by a two-tailed paired t-test on log-transformed TCID 50 /mL titres. ** p = 0.005; *** p = 0.0005.

Article Snippet: Cells were then incubated with 50 μL/well of the primary antibody anti-RVFV nucleoprotein mouse monoclonal antibody (M977, The Native Antigen Company) diluted 1:1000 in blocking buffer, for 1 h at room temperature.

Techniques: Functional Assay, Sequencing, Virus, Luciferase, Activity Assay, Titration, Two Tailed Test, Transformation Assay

Journal: NPJ Vaccines

Article Title: A system to rapidly develop a bunyavirus pseudotyped virus neutralisation assay for pandemic preparedness

doi: 10.1038/s41541-025-01278-8

Figure Lengend Snippet: 100 TCID 50 of RVFV (green) CCHFV (blue) DABV (orange) or OROV (black) VSV-based PV were incubated with serial dilutions of anti-RVFV mAb-144 ( A ), anti-CCHFV mAB-11E7 ( B ), anti-DABV S2A5 ( C ) or anti-OROV ascitic fluid ATCC VR1228AF ( D ), prior to addition to Huh7 cells. Percentage of neutralisation was calculated based on luciferase activity relative to PV only. The graphs show average values ± standard error of the mean of at least two experiments conducted in duplicate.

Article Snippet: Cells were then incubated with 50 μL/well of the primary antibody anti-RVFV nucleoprotein mouse monoclonal antibody (M977, The Native Antigen Company) diluted 1:1000 in blocking buffer, for 1 h at room temperature.

Techniques: Incubation, Luciferase, Activity Assay

Journal: NPJ Vaccines

Article Title: A system to rapidly develop a bunyavirus pseudotyped virus neutralisation assay for pandemic preparedness

doi: 10.1038/s41541-025-01278-8

Figure Lengend Snippet: Plasma from eight donors, seven CCHF convalescent and a negative (−VE) healthy individual, was tested against CCHFV/VSV PV on Huh7 cells. Percentage of neutralisation was calculated based on luciferase activity relative to PV only. The graphs show average values ± standard error of the mean of two experiments conducted in duplicate. Samples were ranked from the highest (top) neutraliser to the lowest ( A ). 60 serum samples from Kenya were tested in a microneutralisation assay using RVFV ZH501 and in a RVFV (ZH548)/VSV PV on Vero cells. For the 41 positive samples, neutralisation titres expressed as 50% neutralisation titre (NT 50 ) were calculated using a 4-parameter regression analysis and log 2 -transformed data. Titres were compared through Deming regression ( B ) and Bland-Altman model ( C ) using GraphPad Prism version 10.3.1.

Article Snippet: Cells were then incubated with 50 μL/well of the primary antibody anti-RVFV nucleoprotein mouse monoclonal antibody (M977, The Native Antigen Company) diluted 1:1000 in blocking buffer, for 1 h at room temperature.

Techniques: Clinical Proteomics, Luciferase, Activity Assay, Transformation Assay