Journal: Cancer Research

Article Title: A Novel Small Molecule Inhibitor of Signal Transducers and Activators of Transcription 3 Reverses Immune Tolerance in Malignant Glioma Patients

doi: 10.1158/0008-5472.can-07-1243

Figure Lengend Snippet: Figure 4. WP1066 enhances phosphorylation of intracellular signaling molecules. Monocytes and Tcells isolated from the PBMCs of GBM patients were incubated with either the medium or medium supplemented with 5 Amol/L WP1066. After 2 h, monocytes and T cells were stimulated for 5 min with 2 Ag/mL LPS and 5 Ag/mL anti-CD3 antibody, respectively. Subsequently, cells were lysed, electrophoretically fractionated in 8% SDS-polyacrylamide gels, transferred to nitrocellulose membranes, and immunoblotted with anti-phosphotyrosine monoclonal antibody 4G10. A, for monocyte membranes, subsequently the same membrane was stripped and reblotted with antibodies to phosphorylated p72Syk (Tyr352), Syk, phosphorylated Hck, Hck, and Lyn. B, for the T-cell membranes, after stripping, the membrane was reblotted with antibodies to phosphorylated ZAP-70 (Tyr319), ZAP-70, p56-Lck, and h-actin.

Article Snippet: T cells from the same patients were isolated with human CD3 microbeads and a magnetic separation column (LS MACS column, Miltenyi Biotec) with the exception of one patient who lacked sufficient PBMCs for both monocyte and T-cell isolations.

Techniques: Phospho-proteomics, Isolation, Incubation, Membrane, Stripping Membranes

Journal: The Journal of Endocrinology

Article Title: ERα expression in T lymphocytes is dispensable for estrogenic effects in bone

doi: 10.1530/JOE-18-0183

Figure Lengend Snippet: Specific inactivation of ERα mRNA expression in T lymphocytes. Two- to seven-month-old gonadal intact female mice were used to study ERα mRNA expression in CD3-positive cells (T lymphocytes) from thymus, muscle, hypothalamus, bone marrow, cortical bone and gonadal fat. Values are given as mean ± s.e.m. ( n = 6–13). *** P < 0.001, Student’s t test, Lck-ERα −/− vs controls.

Article Snippet: CD3-positive cells were extracted from freshly dissected thymus with a mouse CD3ε MicroBead Kit (MACS, Miltenyi Biotec) according to the manufacturer’s protocol.

Techniques: Expressing