Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Flicr , a long noncoding RNA, modulates Foxp3 expression and autoimmunity

doi: 10.1073/pnas.1700946114

Figure Lengend Snippet: Flicr is expressed specifically in Tregs. Flicr expression in various contexts. (A and B) ImmGen mouse immunocyte (39) or GNF mouse organ microarray compendia (40) (dashed lines: background level). (C) During thymic Treg differentiation. (Left) ImmGen data. (Right) RNA-Seq analysis of CD4+ SP thymocytes sorted as shown; y-axis, transcripts per million (tpm). (D) Tissue Tregs. Each point is an individual mouse. Sp, spleen; Col, colonic lamina propria; Panc, pancreas; Adip. tissue, visceral adipose tissue; Muscle, injured muscle at 4 d postcardiotoxin injection. (E) After activation in vitro with anti-CD3/CD28 beads (42) (Left) or in vivo (41) (Right). Rest., resting CD44loCD62Lhi; Act., activated CD44hiCD62Llo Tregs; rpkm, reads per kb per million reads. (F) In vitro converted induced Tregs (TCR activation with IL-2 and TGF-β) (43). (G) Ex vivo Helios−RORγ+ colonic peripheral Tregs (44). (H) In Tconvs transduced with Foxp3 or control retrovectors (11). (I) In Treg-like cells of mice with Foxp3 inactivation by Gfp insertion (Foxp3ΔGfp) (45).

Article Snippet: Total CD4 T cells from spleen and subcutaneous lymph nodes were isolated by magnetic negative selection (Dynabeads Untouched Mouse CD4 T-Cell Kit, 11415D; Thermo Fisher Scientific) and activated in vitro in 200 µL of cRMPI for 72 h with anti-CD3/CD28 beads in a 96-well plate (0.1 M cell/well, 1:1 bead:cell ratio; Dynabeads Mouse T-Activator CD3/CD28; Thermo Fisher Scientific) and 50 U/mL IL-2 (recombinant human IL-2, 212–12; Peprotech).

Techniques: Expressing, Microarray, RNA Sequencing Assay, Injection, Activation Assay, In Vitro, In Vivo, Ex Vivo, Transduction

Journal: Cancer cell

Article Title: KRAS-IRF2 axis drives immune suppression and immune therapy resistance in colorectal cancer

doi: 10.1016/j.ccell.2019.02.008

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: T cell suppression assay A T cell suppression assay was performed as we previously described ( Lu et al., 2017a ; Wang et al., 2016 ) using MACS-sorted MDSCs and CFSE (Invitrogen)-labeled MACS-sorted (Miltenyi Biotec, Cat 130–104-075) CD8 + T cells in anti-CD3-and anti-CD28-coated 96-well plates at an MDSC/ T cell ratio of 0:1, 1:1, with 5.0 × 10 5 MDSCs.

Techniques: Control, Labeling, Recombinant, Enzyme-linked Immunosorbent Assay, Isolation, Cell Isolation, Bicinchoninic Acid Protein Assay, cDNA Synthesis, Microarray, shRNA, Plasmid Preparation, Software, Blocking Assay, Transfection, Staining