Journal: iScience

Article Title: ACE2 interaction with cytoplasmic PDZ protein enhances SARS-CoV-2 invasion

doi: 10.1016/j.isci.2021.102770

Figure Lengend Snippet:

Article Snippet: Spike (SARS-CoV-2) Pseudotyped Lentivirus (Luciferase Reporter) , BPS Bioscience , Cat#79942-1.

Techniques: Recombinant, Bicinchoninic Acid Protein Assay, In Situ, Proximity Ligation Assay, Luciferase, Software

Journal: Molecular Neurobiology

Article Title: NFATc4 Knockout Promotes Neuroprotection and Retinal Ganglion Cell Regeneration After Optic Nerve Injury

doi: 10.1007/s12035-024-04129-0

Figure Lengend Snippet: NFATc4 expression and NFAT transcriptional activity following lentiviral transduction. ( A ) Nfatc4 mRNA expression was assessed in primary hippocampal neurons following transduction with either Lenti-GFP-NFATc4 or Lenti-GFP, using real-time PCR. Raw data were normalized to Gapdh endogenous expression and calculated using 2 −ΔΔCt method. Nfatc4 expression level in non-transduced cells was set as 1. The data are presented as means ± SEM, with individual values obtained from n = 4 replicate treatment. ( B ) Primary hippocampal neurons were co-transduced with NFAT dual-reporter lentivirus and Lenti-GFP-NFATc4 (or other viruses as indicated on the graph) on DIV0 and cultured until DIV3. NFAT transcriptional activity was determined in cell lysates by measuring luciferase activity (n = 4). The results are expressed as a fold induction above baseline activity. The data are presented as means ± SEM, with individual values indicated on the graphs. * P < 0.05, *** P < 0.001. ( C ) Lenti-GFP or Lenti-GFP-NFATc4 were intravitreally injected into NFATc4 −/− retinas, followed by NFATc4 staining three weeks later. The retinas were stained using the antibodies indicated in Retina cryosection staining (primary: NFATc4, 1:500, Merck, USA; secondary: anti-rabbit conjugated to Alexa Fluor 488, 1:500, Thermo Fisher, USA). Representative images are presented. Scale bar: 100 μm

Article Snippet: NFAT luciferase reporter lentivirus and firefly luciferase lentivirus were sourced from BPS Bioscience (USA).

Techniques: Expressing, Activity Assay, Transduction, Real-time Polymerase Chain Reaction, Cell Culture, Luciferase, Injection, Staining

Journal: The Journal of Clinical Investigation

Article Title: Loss of ARPC1B impairs cytotoxic T lymphocyte maintenance and cytolytic activity

doi: 10.1172/JCI129388

Figure Lengend Snippet: ( A and B ) Percentage lysis of P815 (after 2 hours) by HD or ARPC1B-deficient patient hCTLs at the indicated effector-to-target ratios in the presence ( A ) or absence ( B ) of IL-2. ( C ) Percentage lysis of P815-NucLight Red targets over time measured by IncuCyte killing assay using HD or ARPC1B-deficient patient hCTLs that have been deprived of IL-2 for 16 hours. ( D ) Western blot analysis of p-ERK1/2 and ERK1/2 expression in HD and ARPC1B-deficient patient hCTLs stimulated for 15 minutes with plate-bound anti-CD3 antibody at the indicated concentrations. Numbers indicate the fold change (ratio) of p-ERK2 expression following stimulation in ARPC1B-deficient patient and HD after normalization to total ERK2 expression. ( E ) Representative flow cytometry plot and quantitation ( F ) of LAMP1-PE (CD107a) uptake in HD and ARPC1B-deficient patient hCTLs (gated on CD8 + cells) after 2 hours incubation under the indicated conditions. ( G and H ) Immunoblot of granzyme B ( G ) and ARPC1A ( H ) in HD and ARPC1B-deficient patient in the presence or absence of IL-2. MW markers in kD. ( A – C ) Data are shown as mean of 3 independent experiments; error bars represent SEM. ( D , G , H ) Data are representative of 4 independent experiments.

Article Snippet: To generate P815-NucLight Red, P815 was transduced with the IncuCyte NucLight Red Lentivirus (4625, Essen Bioscience) to stably label the nuclei with mKate2 red fluorescent protein.

Techniques: Lysis, Western Blot, Expressing, Flow Cytometry, Quantitation Assay, Incubation