Journal: Journal of Nuclear Medicine

Article Title: 225 Ac/ 89 Zr-Labeled N4MU01 Radioimmunoconjugates as Theranostics Against Nectin-4–Positive Triple-Negative Breast Cancer

doi: 10.2967/jnumed.124.268387

Figure Lengend Snippet: Flow cytometry binding, internalization, and radioligand binding assay of anti–nectin-4 antibody (N4MU01) and its conjugates with (A) MDA-MB-231, MCF-7, MDA-MB-468, and 4T1. nectin-4 cells. Dose-dependent binding was observed in nectin-4–expressing cell lines, MDA-MB-468, MCF-7, and 4T1. nectin-4 . No binding was observed in MDA-MB-231 control cell line. (B) Internalization of N4MU01: MDA-MB-468, MCF-7, and MDA-MB-231 cells were treated with Incucyte FabFluor-labeled N4MU01 (4 μg/mL); High-definition phase and red fluorescence images (×10 magnification) were captured every 2 h for 48 h. All data are shown as mean ± SEM ( n = 3), and average internalization values for MCF-7 and MDA-MB-468 were compared with that of MDA-MB-231 using ANOVA. Estimation of K D values using radioligand binding assay for (C) [ 89 Zr]Zr-DFO-N4MU01 in nectin-4–positive MCF-7 and (D) [ 225 Ac]Ac-Macropa-N4MU01 in nectin-4–positive MDA-MB-468 cells. **** P < 0.0001. FITC-A = fluorescein isothiocyanate area; FL1-A = fluorescence intensity from channel 1 (green) area; FL2-A = fluorescence intensity from channel 2 (orange) area; PE-A = phycoerythrin area.

Article Snippet: The in vitro cytotoxicity (inhibitory concentration of 50% [IC 50 ]) of [ 225 Ac]Ac-Macropa-N4MU01, control immunoconjugates anti-CD20 [ 225 Ac]Ac-Macropa-rituximab, and unlabeled N4MU01 in nectin-4–positive MDA-MB-468, MCF-7, and nectin-4–negative MDA-MB-231 was determined using Incucyte Cytotox Red reagent in an Incucyte S3 live-cell imager (Essen BioScience) as previously reported (details in the supplemental materials) ( ).

Techniques: Flow Cytometry, Binding Assay, Radio Ligand Binding Assay, Expressing, Control, Labeling, Fluorescence

Journal: Cell Reports

Article Title: Quantitative Temporal Proteomic Analysis of Vaccinia Virus Infection Reveals Regulation of Histone Deacetylases by an Interferon Antagonist

doi: 10.1016/j.celrep.2019.04.042

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used at the indicated dilution, in blocking solution: rabbit anti-IFIT1 (1:1000, cat. no. PA3-848, Thermo Fisher Scientific), mouse anti-IFITM1/2/3 (1:500, cat. no. sc-374026, Santa Cruz Biotechnology), mouse anti-COL6A2 (1:500, cat. no. sc-374566, Santa Cruz Biotechnology), mouse anti-COL1A2 (1:500, cat. no. sc-376350, Santa Cruz Biotechnology), mouse anti-TRIM5 (1:500, cat. no. SC-373864, Santa Cruz Biotechnology), mouse anti-HDAC5 (1:500, cat. no. sc-133225, Santa Cruz Biotechnology), mouse anti-HDAC1 (1:500, cat. no. sc-81598, Santa Cruz Biotechnology), mouse anti-FLAG, (1:1000, F3165, Sigma-Aldrich), mouse anti-Myc (1:1000, cat. no. 2276, Cell Signaling Technology), rat anti-α-tubulin (1:10,000, clone YL1/2, cat. no. MCA77G, Serotec), rabbit anti-C6 (1:1000, described in , mouse anti-D8 (1:1000, described in ( )).

Techniques: Virus, Western Blot, Derivative Assay, Subcloning, Recombinant, Protease Inhibitor, Electron Microscopy, Bicinchoninic Acid Protein Assay, TA Cloning, Sequencing, Mass Spectrometry, CRISPR, Disruption, Plasmid Preparation, Software, Quantitative Proteomics, Imaging, Flow Cytometry, Fluorescence, Microscopy