Journal: Nanomaterials

Article Title: Cytochalasin B Treatment and Osmotic Pressure Enhance the Production of Extracellular Vesicles (EVs) with Improved Drug Loading Capacity

doi: 10.3390/nano12010003

Figure Lengend Snippet: The proteins expressions, cellular uptake, and dendrimer loading capability of hypo-CIMVs compared to naturally-secreted EVs, hypo-EVs, and CIMVs: ( A ) Amount of total vesicular protein ( n = 3); ( B ) Expression of transmembrane protein CD81 compared to intravesicular protein MIF; ( C ) Immunogold TEM images for assessing the CD63 (transmembrane marker) expression; ( D ) Cellular uptake of the produced EVs; ( E – G ) Loading of ( E ) G7-NH2, ( F ) G7-COOH, and ( G ) G7-Ac to the produced EVs. Significance levels are indicated as # p < 0.10, * p < 0.05, and ** p < 0.01, compared to the EVs obtained from non-CB treated cells. The dendrimer loading to the EVs was analyzed from at least three independent experiments ( n ≥ 3).

Article Snippet: The blot was blocked with 5% skim milk for 1 h followed by overnight incubation in primary antibody against CD81 (monoclonal anti-CD81 SC-166028, Santa Cruz Biotechnology, San Jose, CA, USA) and macrophage migration inhibitory factor (MIF) (monoclonal anti-MIF, MAB289, R&D Systems) at 4 °C.

Techniques: Expressing, Marker, Produced

Journal: Journal for Immunotherapy of Cancer

Article Title: Intratumor expanded T cell clones can be non-sentinel lymph node derived in breast cancer revealed by single-cell immune profiling

doi: 10.1136/jitc-2021-003325

Figure Lengend Snippet: Characteristics of T cells residing in primary breast cancers and paired sentinel LNs. (A) UMAP of T cells with color-coded clusters. (B) Velocities (arrows) of T cells in the different clusters. (C) Expression of characteristics genes in each T cell subset. (D) Diffusion map of all CD8 +T cells. (E) Average expression of naïve, cytolytic and exhaustion markers across diffusion component one and diffusion component 2. (F) Diffusion map of cytolytic (FGFBP2), exhaustion (HAVCR2) and naïve (LEF1) markers. (G) Scatter plots of T cells in the space of activation and exhaustion scores color-coded by expression levels of FGFBP2, HAVCR2 and LEF1. DC, dendritic cell; FGFBP2, fibroblast growth factor binding protein 2; HAVCR2, hepatitis A virus cellular receptor 2; LEF1, lymphoid enhancer binding factor 1; LN, lymph node; UMAP, uniform manifold approximation and projection.

Article Snippet: Multiplex staining was performed using the Opal 4-Color Manual IHC Kit (NEL810001KT) with the anti-CD3 (1:150; ZA-0508, ZSGB-BIO), anti-CD20 (1:100; ZM-0039, ZSGB-BIO), anti-CCL4 (1:50, PA5-23681, ThermoFisher), anti-CD8 (1:100, ab4055, abcam), anti-PNAD (1:100, #120801, Biolegend), anti-PD1(1:100, ab52587, Abcam) and anti-CD103 (1:100, ab254017, Abcam) antibodies with subsequent visualization with fluorescein AF-690 (1:75), AF-520 (1:75) and AF-570 (1:50).

Techniques: Expressing, Diffusion-based Assay, Activation Assay, Binding Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Intratumor expanded T cell clones can be non-sentinel lymph node derived in breast cancer revealed by single-cell immune profiling

doi: 10.1136/jitc-2021-003325

Figure Lengend Snippet: The TLS in primary breast cancer may induce intratumoral novel T cell clones. (A) DEGs between expanded clonal CD8 T cells and antigen-experienced non-clonal CD8 T cells. Significant DEGs were screened by p<0.05 and avg_FC>=1.5. (B) Multiplex immunofluorescence imaging of tumor tissues. Red - CD3+, white - CCL4+. CD3 +CCL4+ is the signature of clonal effector T cells. Green - CD20 +B cells. The aggregation of T and B cells represents TLS. CA, cancer; DEGs, differentially expressed genes; TLS, tertiary lymphoid structures.

Article Snippet: Multiplex staining was performed using the Opal 4-Color Manual IHC Kit (NEL810001KT) with the anti-CD3 (1:150; ZA-0508, ZSGB-BIO), anti-CD20 (1:100; ZM-0039, ZSGB-BIO), anti-CCL4 (1:50, PA5-23681, ThermoFisher), anti-CD8 (1:100, ab4055, abcam), anti-PNAD (1:100, #120801, Biolegend), anti-PD1(1:100, ab52587, Abcam) and anti-CD103 (1:100, ab254017, Abcam) antibodies with subsequent visualization with fluorescein AF-690 (1:75), AF-520 (1:75) and AF-570 (1:50).

Techniques: Clone Assay, Multiplex Assay, Immunofluorescence, Imaging

Journal: eLife

Article Title: Binary outcomes of enhancer activity underlie stable random monoallelic expression

doi: 10.7554/eLife.74204

Figure Lengend Snippet:

Article Snippet: Antibody , (mouse monoclonal) anti-CD8.1 (clone 116–13.1) , BioXCell , RRID: AB_10949065 Catalog # BE0118 , FACS (1:250).

Techniques: Generated, Microinjection, Electroporation, CRISPR, Purification, cDNA Synthesis, Cell Isolation, Multiplex Assay, TA Cloning, Subcloning, Staining, Flow Cytometry, Recombinant, Cell Culture, Software