Journal: Cell reports

Article Title: Macrophage Exosomes Resolve Atherosclerosis by Regulating Hematopoiesis and Inflammation via MicroRNA Cargo

doi: 10.1016/j.celrep.2020.107881

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-Mouse Macrophages / Monocytes Purified (Clone MOMA-2) (rat IgG2b) , Cedarlane , Cat# CL89154; RRID: AB_10086520.

Techniques: Western Blot, Binding Assay, Flow Cytometry, Purification, Recombinant, SYBR Green Assay, Red Blood Cell Lysis, Reverse Transcription, Blocking Assay, Plasmid Preparation, Sterility, Transfection, Bicinchoninic Acid Protein Assay, Subcloning, RNA Sequencing, Qubit Protein Assay, Cell Culture, Software

Journal: PLoS Genetics

Article Title: Pregnancy-Induced Noncoding RNA ( PINC ) Associates with Polycomb Repressive Complex 2 and Regulates Mammary Epithelial Differentiation

doi: 10.1371/journal.pgen.1002840

Figure Lengend Snippet: (A–C) RNA was isolated from FACs sorted mammary populations including, virgin luminal (VL) and basal (VB) as well as pregnant luminal (PL) and basal (PB). (A) qPCR showed Krt8 expression was enriched in the luminal populations (VL and PL), (B) Krt14 was enriched in the basal populations (VB and PB) and (C) mPINC was enriched in the pregnant luminal population, (D–F) MECs were FACs sorted into luminal and basal populations using CD24 and CD29. The luminal population was selected and further sorted into mature luminal (ML), luminal progenitors (LP), and alveolar progenitors (AP) using CD14 and ckit. (D) FACs dot plots showing CD24 and CD29 (left panel) as well as CD14 and ckit (right panel) from virgin MECs. (E) qPCR showed Ly6a and Wnt4 enriched in the ML population and Elf5 in the LP population thus verifying the purity of each population. (F) mPINC was enriched in the luminal progenitors and alveolar progenitors. Data represent mean ±SD (n = 3). Target genes were normalized to Gapdh .

Article Snippet: Mammary epithelial cells were subsequently resuspended at a density of 1×10 7 cells/ml and stained with anti-mouse CD24 PE (Stem Cell Technologies, 1∶100), anti-CD49f FITC (Stem Cell Technologies, 1∶100), anti-mouse CD24 APC (Biolegend, 1∶100), anti-mouse CD29 Pacific Blue (BioLegend, 1∶100), anti-mouse CD14 FITC (eBiosiences, 1∶80) and anti-mouse ckit PE (Clone ACK4, Cedarlane Laboratories, 1∶50).

Techniques: Isolation, Expressing

Journal:

Article Title: E2F1 and E2F2 Determine Thresholds for Antigen-Induced T-Cell Proliferation and Suppress Tumorigenesis

doi: 10.1128/MCB.21.24.8547-8564.2001

Figure Lengend Snippet: Loss of E2F1 and E2F2 decreases thresholds for antigen-induced T-cell proliferation. (A) Lymphocytes from 6-week-old male E2F1+/− E2F2+/− and DKO littermates were isolated from the lymph nodes and cultured with 2 μM OVA in RP10 medium. During the indicated time windows, BrdU was added, and the cells were harvested at the end of the window. BrdU incorporation was detected by flow cytometry. We found that 5.5% of E2F1+/− E2F2+/− and 4.9% of DKO lymphocytes cultured without antigen and labeled with BrdU from 8 to 42 h were positive for BrdU incorporation. (B) Lymphocytes from lymph nodes of DO11.10 TCR transgenic 6-week-old female littermates of the indicated genotypes were harvested and cultured with the indicated concentrations of antigenic peptide (OVA) and 1 μM BrdU for 36 h. BrdU incorporation was detected by flow cytometry. The y axis represents cell number. The percentages of cells that were positive for BrdU incorporation are indicated. (C) Lymphocytes from the spleens of 6.5-week-old female mice of the indicated genotypes were harvested, stained with CFSE, and cultured with OVA at the indicated concentrations for 72 h. The DKO and E2F1+/− E2F2+/− mice were littermates, and the WT mouse was female and age matched. The number of cell divisions (as indicated for divisions 1 to 5 above plots; P, parental) in T cells was determined by the intensity of CFSE. The position of the parental (P) peak is based on the analysis of the same cells cultured without antigen, as shown in Fig. ​Fig.4B.4B. The ∗ peak reflects nonlymphocytes that contaminated the T-cell gate. (D) Lymphocytes from DO transgenic littermates of the indicated genotypes cultured with 0.2 μM OVA for 24 h (or unstimulated; 0 h) were stained with fluorochrome-linked anti-Thy1.2, anti-CD25, and anti-CD69 antibodies. The expression of CD69 and CD25 was determined in T cells (Thy1.2+ lymphocytes) by flow cytometry, and the percentages of cells that upregulated both CD69 and CD25 expression are indicated.

Article Snippet: The following Pharmingen antibodies were used: phycoerythrin (PE)-linked α-CD4, Cy-Chrome-α-CD8, PE-α-CD25, biotin-α-CD25 (together with streptavidin-Cy-Chrome), allophycocyanin (APC)-linked α-B220, APC-α-Thy1.2, APC-α-GR-1, APC-α-Ter119, APC-α-CD34, PE-α-CD43, fluorescein isothiocyanate (FITC)-linked TCR variable-chain beta (Vβ) 5, FITC-TCR Vβ6, and FITC-CD44.

Techniques: Isolation, Cell Culture, BrdU Incorporation Assay, Flow Cytometry, Labeling, Transgenic Assay, Staining, Expressing