Journal: Immunity

Article Title: The Chromatin-Modifying Enzyme Ezh2 Is Critical for the Maintenance of Regulatory T Cell Identity after Activation

doi: 10.1016/j.immuni.2015.01.007

Figure Lengend Snippet: (A) Microarray analysis to reveal CD28-dependent genes by comparison of human naive CD4+ T cells that were unstimulated, stimulated with anti-CD3, or stimulated with anti-CD3 and anti-CD28 antibodies for 24 hr. Heat map of known histone lysine demethylases, methyltransfereses, acetyltransferases, and deacetylases that were significantly differentially expressed by comparing anti-CD3+CD28-stimulated versus unstimulated and anti-CD3+ CD28-stimulated versus anti-CD3 stimulated (36 out of all 9,824 DEGs, FDR < 0.05) are plotted.

Article Snippet: Cells were activated with anti-CD3 and anti-CD28 coated beads (Dynabeads Mouse T-Activator CD3/CD28, Invitrogen) at a ratio of 1:1 (cell:bead), beads coated selectively with anti-CD3 or anti-CD3 and anti-CD28 (T Cell Activation/Expansion Kit mouse, Miltenyi Biotec), or 1 μg/ml soluble antibodies against CD3 (clone 145-2C11) ±CD28 (clone PV-1).

Techniques: Microarray

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-7R-Dependent Phosphatidylinositol 3-Kinase Competes with the STAT5 Signal to Modulate T Cell Development and Homeostasis.

doi: 10.4049/jimmunol.1900456

Figure Lengend Snippet: FIGURE 6. Survival and homeostatic proliferation of T cells are impaired in IL-7R–Y449F mice but enhanced in IL-7R–M452L mice. (A) Cell numbers of CD4 T, CD8 T, iNKT, Treg, and gd T cells in lymph nodes of WT, IL-7R–Y449F, and IL-7R–M452L mice (n = 8). (B) Flow cytometry analysis of Bcl-2 expression in naive CD4 and CD8 T cells. Gray shaded curves, isotype-matched control Ab. (C) qPCR analysis of Bcl-2 mRNA on naive CD4 and CD8 T cells (n = 5). (D) Naive CD4 T cells were isolated from lymph nodes and cultured with 50 ng/ml IL-7 for 4 d. Proliferation of naive CD4 T cells was assessed by CFSE dilution assay. CFSE+ cells were divided into three fractions: CFSEhigh (no division), CFSEintermediate (one or two divisions), and CFSElow

Article Snippet: Isolated naive CD4 and CD8 T cells were labeled with 2 mM CFSE (Dojindo Laboratories) in PBS at 37 ̊C for 10 min. For in vitro proliferation and activation, cells were cultured with 50 ng/ml IL-7, or with 4 mg/ml plate-bound anti-CD3 Ab, and 4 mg/ml soluble anti-CD28 Ab (PV-1).

Techniques: Flow Cytometry, Expressing, Control, Isolation, Cell Culture, Dilution Assay

Journal: Frontiers in Immunology

Article Title: FMNL1 and mDia1 promote efficient T cell migration through complex environments via distinct mechanisms

doi: 10.3389/fimmu.2024.1467415

Figure Lengend Snippet: Key resources.

Article Snippet: Polyclonal T cells were harvested from C57BL/6 mouse superficial and mesenteric lymph nodes and activated using plate-bound anti-CD3 (2C11, BioXCell), soluble anti-CD28 (PV-1, BioXCell), and autologous splenocytes for two days.

Techniques: Recombinant, Fractionation, Cell Culture