Journal: bioRxiv

Article Title: A Rapid Bead-Based Assay For Screening Of SARS-CoV-2 Neutralising Antibodies

doi: 10.1101/2021.10.13.464050

Figure Lengend Snippet: a: GFP nanobody with GST tag immobilized on Sepharose beads were incubated with HEK 293T ACE2-GFP lysate b: HEK 293T cells stably expressing SARS-CoV-2 S1-OFP c: whole-cell extract from HEK293T S1-OFP stable cells readily bind to the ACE2-GFP Beads d: whole-cell extract from HEK293T-S1-OFP cells at a concentration of 100 ug/ml incubated with RFP Trap beads for indicated time points, Respective overlay and RFP channels are shown e: Quantification of the S1-OFP fluorescence intensity at indicated time points are shown f: The S1-OFP beads incubated with whole cell lysate from HEK293T-ACE2-GFP cells at a concentration of 100ug/ml for indicated time points, respective overlay, GFP channel and RFP channels are shown g: Quantification of the ACE2-GFP fluorescence intensity at indicated time points are shown h: The S1-OFP beads were pre-incubated with the indicated dilution of neutralizing antibody for 1 hour followed by ACE2 GFP extract, respective overlay, GFP channel, and RFP channels are shown i: Quantification of the ACE2-GFP fluorescence intensity at indicated dilutions of neutralizing antibody are shown. p-value calculated using Two-way ANOVA with Tukey’s Post hoc hypothesis (0.0001>***,0.001>**, 0.01>*)

Article Snippet: GFP nanobody and mCherry nanobody prokaryotic expression vectors were procured from Addgene (#49172, #70696).

Techniques: Incubation, Stable Transfection, Expressing, Concentration Assay, Fluorescence

Journal: Nature communications

Article Title: Virus-modified paraspeckle-like condensates are hubs for viral RNA processing and their formation drives genomic instability.

doi: 10.1038/s41467-024-54592-5

Figure Lengend Snippet: Fig. 3 | The viral protein ORF11 associates with v-mPS. A Graph of proteins over a 10% abundance ratio and 4% abundance change in the SFPQ TMT-MS at 24 h (n = 2). ORF11 is highlighted in orange, proteins involved in RNA processing are highlighted purple and DEAD/DEAH box helicases and hnRNP proteins are blue. Proteins con- firmed via IF as co-localising are marked with arrows. B Western blot analysis of FLAG Co-IPs in TREx-BCBL1-RTA FLAG-ORF11 O/E cells during latent or 24 h post- lytic replication and probed with antibodies against FLAG (ORF11), SFPQ, ORF57 and GAPDH (n = 3). C Western blot analysis of FLAG Co-IPs in TREx-BCBL1-RTA

Article Snippet: Protein purification Wildtype ORF11 cloned into popinJ vector (Addgene: #26045) was transformed into Lemo21(DE3) competent cells and innoculated in 1 L of ampicillin selective LB broth at 37 °C with 180 RPM shaking until OD600 reached 0.6 nm absorbance.

Techniques: Western Blot