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

Article Title: Expansion of bone marrow IFN-alpha-producing dendritic cells in New Zealand Black (NZB) mice: high level expression of TLR9 and secretion of IFN-alpha in NZB bone marrow.

doi: 10.4049/jimmunol.173.8.5283

Figure Lengend Snippet: FIGURE 2. Identification of NZB IPDC subsets. A, Freshly isolated BM, spleen, LN, liver, thymus, and lung lymphocytes were resolved by staining for NK1.1, CD11c, B220, and CD4. The NK1.1CD11c cell population can be resolved into two subpopulations, CD4 IPDC and CD4 IPDC cells. B, Levels of mRNA transcripts for TdT, 0, mb-1, B29, and the internal control gene -actin in the NZB two BM IPDC subsets were analyzed by RT-PCR. Primers used are listed in Table I. Our negative controls include lymphoid and myeloid DC cells. C, Morphology of the NZB BM two IPDC subsets. Cytospin preparations of sorted BM IPDCs were stained with May-Giemsa and photographed at 100. Upper panel, Shows freshly isolated and uncultured cells. Lower panel, Shows IPDC cells cultured for 48 h with type II CpG ODN 2006 (2 M). D, The comparison of the surface phenotypes of BM IPDC subsets between NZB and BALB/c mice. Isolated BM cells were stained with CD4-FITC, CD11c-PE, and B220-APC, followed by staining with biotin-labeled Abs for CD80, CD86, MHC class II, or CD45RA, and then PE Cy5.5-conju- gated streptavidin.

Article Snippet: The following unconjugated or directly conjugated mAbs were used: purified anti-CD16/CD32 (Fc II/IIIR, 93) (e-Bioscience, San Diego, CA); FITC-labeled anti-CD4 (GK1.4), CD11b (M1/70), and CD43 (S7) (BD Pharmingen, San Diego, CA); MHC class II (M5/114.15.2) (Miltenyi Biotec, Auburn, CA); PE-labeled antiCD11c (HL3) (BD Pharmingen) and CD127 (IL-7R, A7R34) (e-Bioscience); biotin-labeled anti-CD4 (GK1.4), CD11b (Mac-1, M1/70), CD19 (1D3), CD24 (heat-stable Ag (HSA), M1/69), CD16/32 (Fc RIII/II, 2.4G2), CD40 (3/23), CD80 (B7.1, 16-10A1), CD86 (B7.2, GL1), CD117 (c-kit, 2B8), CD123 (IL-3R, 5B11), TCR -chain (H28-710), and Ly-6G (Gr-1, RB6-8C5) (BD Pharmingen); NK1.1 (PK136) (e-Bioscience); and allophycocyanin-labeled anti-B220 (B220, RA3-6B2) (Caltag Laboratories).

Techniques: Isolation, Staining, Control, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Comparison, Labeling

Journal: Cell Death & Disease

Article Title: Kaiso depletion attenuates the growth and survival of triple negative breast cancer cells

doi: 10.1038/cddis.2017.92

Figure Lengend Snippet: Schematic diagram of proposed model for Kaiso's role in TNBC. ( a ) Kaiso interacts with both wt p53 and mutant p53 in BCa cells and this differential interaction may modulate Kaiso's function in apoptosis. ( b ) In TNBC cells lacking wt p53 (but expressing mutant p53), Kaiso might directly or indirectly inhibit the activation of the pro-apoptotic genes Bax and PUMA, which leads to tumor survival. However, Kaiso's inhibitory effect on Bax protein expression may be attenuated by Kaiso interaction with other proteins like p120 ctn . Kaiso may also activate c-Myc, Cyclin D1 and BRCA1 expression independently or in collaboration with other cofactors in TNBC cells, which would also promote TNBC cell growth and survival

Article Snippet: Overnight incubations were performed at 4 °C using the following primary antibodies at their respective dilutions; Kaiso-specific rabbit polyclonal (gift from Dr. Reynolds; 1:5000), mouse monoclonal antibody against c-Myc (SantaCruz (9E10); 1:500), rabbit polyclonal antibody against Cyclin D1 (US Biological (144418); 1:5000), p120 ctn -15D2 specific mouse monoclonal (gift from Dr. Reynolds; 1:1000 ), Bax-specific rabbit monoclonal (1:500; CST-5023), PUMA-specific rabbit monoclonal (1:500; CST-12450), p53-specific rabbit polyclonal (1:2000; Abcam-ab32049), cleaved PARP-specific rabbit monoclonal (1:1000; CST-5625), BRCA1-specific rabbit polyclonal (1:2000; Abcam-ab131360) and mouse anti- β -actin monoclonal (1:50 000; Sigma Aldrich).

Techniques: Mutagenesis, Expressing, Activation Assay