Journal: Cell reports

Article Title: Human papillomavirus E5 suppresses immunity via inhibition of the immunoproteasome and STING pathway

doi: 10.1016/j.celrep.2023.112508

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit monoclonal anti-phospho-IRF3 (Ser396) (clone 4D4G) , Cell Signaling Technology , Cat#: 4947; RRID: AB_823547.

Techniques: Recombinant, cDNA Synthesis, SYBR Green Assay, Protease Inhibitor, Suspension, Activity Assay, Reporter Assay, Plasmid Preparation, Expressing, Software

Journal: PLoS ONE

Article Title: Recombinant Neuregulin 1 Does Not Activate Cardiomyocyte DNA Synthesis in Normal or Infarcted Adult Mice

doi: 10.1371/journal.pone.0115871

Figure Lengend Snippet: (A) Use of BrdU to monitor cardiomyocyte DNA synthesis in non-injured adult mice receiving 9 consecutive daily injections of NRG1β1 (BrdU was delivered using a mini-osmotic pump). Left panel shows anti-β-galactosidase immune reactivity, middle panel shows anti-BrdU immune reactivity, and right panel shows the merged image. Arrow indicates a BrdU positive cardiomyocyte nucleus, arrowhead indicates a BrdU positive non-cardiomyocyte nucleus. Bar = 10 microns. (B) BrdU incorporation in the nuclei of the small intestine microvilli epithelial cells of an NRG1β1-treated mouse. Note the absence of BrdU signal in the muscularis mucosae zone (asterisk). Bar = 10 microns. (C) Use of 3 H-Thy to monitor cardiomyocyte DNA synthesis in non-injured adult mice receiving 9 consecutive daily injections of NRG1β1 ( 3 H-Thy was delivered as a single bolus 1 hour after the last NRG1β1 treatment). Arrow indicates a 3 H-Thy positive cardiomyocyte nucleus, arrowhead indicates a 3 H-Thy positive non-cardiomyocyte nucleus. Bar = 10 microns.

Article Snippet: Experimental mice were treated with recombinant human NRG1β1 (corresponding to the EGF domain, amino acid residues 176–256, #396-HB, R&D Systems, Minneapolis, MN), at a dose of 2.5 micrograms per mouse per IP injection, dissolved in saline containing 0.1% Bovine Serum Albumin (BSA); control mice received vehicle alone.

Techniques: DNA Synthesis, BrdU Incorporation Assay

Journal: PLoS ONE

Article Title: Recombinant Neuregulin 1 Does Not Activate Cardiomyocyte DNA Synthesis in Normal or Infarcted Adult Mice

doi: 10.1371/journal.pone.0115871

Figure Lengend Snippet: Cardiomyocyte DNA synthesis in adult MHC-nLAC mice following vehicle or NRG1β1 injection.

Article Snippet: Experimental mice were treated with recombinant human NRG1β1 (corresponding to the EGF domain, amino acid residues 176–256, #396-HB, R&D Systems, Minneapolis, MN), at a dose of 2.5 micrograms per mouse per IP injection, dissolved in saline containing 0.1% Bovine Serum Albumin (BSA); control mice received vehicle alone.

Techniques: DNA Synthesis, Injection, Control

Journal: PLoS ONE

Article Title: Recombinant Neuregulin 1 Does Not Activate Cardiomyocyte DNA Synthesis in Normal or Infarcted Adult Mice

doi: 10.1371/journal.pone.0115871

Figure Lengend Snippet: (A) Western blot demonstrating the levels of total Erk1/2 p42/p44, P-Erk1/2[Thr 202 /Thy 204 ] and RLC in mice treated with NRG1β1 or vehicle (hearts harvested and processed 90 minutes after treatment). Densometric quantitation revealed that NRG1β1 treatment resulted in a 987% increase in the level of ERK1 phosphorylation, a 5727% increase in the level of ERK2 phosphorylation, and a 21% increase in the level of phosphorylated RLC vs. vehicle-treated mice (p<0.01, Student’s t-test). (B) Non-cardiomyocyte 3 H-Thy nuclear labeling index in non-injured adult mice following 9 consecutive daily injections of NRG1β1 (5 sections analyzed from each of 4 independent mice) or vehicle (4 sections analyzed from each of 4 independent mice). *: p<0.05 vs. vehicle treated animals, Student’s t-test.

Article Snippet: Experimental mice were treated with recombinant human NRG1β1 (corresponding to the EGF domain, amino acid residues 176–256, #396-HB, R&D Systems, Minneapolis, MN), at a dose of 2.5 micrograms per mouse per IP injection, dissolved in saline containing 0.1% Bovine Serum Albumin (BSA); control mice received vehicle alone.

Techniques: Western Blot, Quantitation Assay, Phospho-proteomics, Labeling

Journal: Foods

Article Title: Preparation, Biological Activities, and Potential Applications of Hen Egg-Derived Peptides: A Review

doi: 10.3390/foods13060885

Figure Lengend Snippet: Preparation methods of HEPs.

Article Snippet: , Chemical synthesis , Egg white protein , AAPPTEC 396 Automated Peptide Synthesizer. The 9-Fluorenylmethoxycarbonyloxy (Fmoc)-protected amino acid synthesis method peptide (RVPSLM). , α-glucosidase inhibitory activity , IC 50 23.07 μmol/L , [ ] .

Techniques: Membrane, Activity Assay, Recombinant, Foaming, Sonication, Control, De-Phosphorylation Assay, Antioxidant Activity Assay, Incubation, Concentration Assay, Emulsification

Journal: iScience

Article Title: Aberrant STING activation promotes macrophage senescence by suppressing autophagy in vascular aging from diabetes

doi: 10.1016/j.isci.2024.111594

Figure Lengend Snippet:

Article Snippet: Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb , Cell Signaling Technology , Cat#4947; RRID: AB_823547.

Techniques: Recombinant, Cell Isolation, Staining, ROS Assay, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, SYBR Green Assay, Plasmid Preparation, Software, Microscopy, Transmission Assay, Gene Expression

Journal: Cell Reports

Article Title: SARS-CoV-2 ORF9b inhibits RIG-I-MAVS antiviral signaling by interrupting K63-linked ubiquitination of NEMO

doi: 10.1016/j.celrep.2021.108761

Figure Lengend Snippet: The N-terminus of ORF9b mediates its interaction with NEMO upon viral infection (A–C) Inhibitory effects of ORF9b on the activation of IFN-β/NF-κB/IRF3 promoters by SeV and the RIG-I-MAVS signaling components. Similar to Figure 1 C, except that the luciferase reporter activities were induced by SeV infection for 12 h or by transfection of RIG-I(N)-, MAVS-, TBK1-, IRF3(S396D)-, and IKKβ-expressing vectors into HEK293T cells for 24 h. IFN-β-Luc (A), NF-κB-Luc (B), and IRF3-Luc (C) reporter activities are normalized to that of Renilla luciferase and shown as fold induction. Data are represented as means ± SDs calculated from three independent experiments ( ∗ p < 0.05, ∗∗ p < 0.01, N.S., non-significant; t test). See also Figure S3 A. (D) Co-immunoprecipitation (coIP) determining the interaction between ORF9b and the RIG-I-MAVS signaling components. HEK293T cells were transfected with plasmid encoding HA-ORF9b-GFP, together with various expressing vectors for the FLAG-tagged RIG-I-MAVS signaling components as indicated. At 24 h post-transfection, cells were infected with or without VSV for 12 h. Immunoprecipitation was conducted using anti-hemagglutinin (HA) beads. See also Figure S3 B. (E) Cellular colocalization of ORF9b and endogenous NEMO. NCI-H1299 cells were co-transfected with expressing vector for HA-ORF9b-GFP for 24 h and were mock-infected or infected with SeV for another 12 h. After immunofluorescent staining of cells with anti-NEMO and Alexa Fluor 594-conjugated secondary antibodies, fluorescent images were taken. Nucleus was labeled with DAPI. Scale bar, 10 μm. See also Figure S3 C. (F) CoIP mapping the motif of ORF9b that is essential for its interaction with NEMO. Plasmid encoding FLAG-NEMO was transfected into HEK293T cells together with various vectors expressing HA-tagged ORF9b and its mutants as indicated. At 24 h post-infection, cells were mock infected or infected with VSV for 12 h. Immunoprecipitation was conducted using anti-FLAG beads. See also D and S3E. (G) Immunoprecipitation of ORF9b with endogenous NEMO under SARS-CoV-2 RNA stimulation. HPAEpiC were transfected with expressing vectors for FLAG-tagged ORF9b or ORF9bΔN30 for 24 h and were then stimulated with SARS-CoV-2 RNA for the indicated time. Immunoprecipitation using anti-flag beads and subsequent immunoblotting was conducted to examine the endogenous NEMO protein levels in each individual sample.

Article Snippet: Phospho-IRF3 (Ser396) Rabbit mAb , Cell Signaling Technology , Cat# 29047.

Techniques: Infection, Activation Assay, Luciferase, Transfection, Expressing, Immunoprecipitation, Plasmid Preparation, Staining, Labeling, Western Blot

Journal: Cell Reports

Article Title: SARS-CoV-2 ORF9b inhibits RIG-I-MAVS antiviral signaling by interrupting K63-linked ubiquitination of NEMO

doi: 10.1016/j.celrep.2021.108761

Figure Lengend Snippet:

Article Snippet: Phospho-IRF3 (Ser396) Rabbit mAb , Cell Signaling Technology , Cat# 29047.

Techniques: Virus, Recombinant, Modification, Saline, Protease Inhibitor, Reporter Assay, Extraction, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Sequencing, Software