Journal:

Article Title: Antigen-pulsed dendritic cells expressing macrophage-derived chemokine elicit Th2 responses and promote specific humoral immunity

doi:

Figure Lengend Snippet: Chemotaxis of human and murine T lymphocytes to conditioned media of genetically modified DCs. (a) CEM cells. Human T lymphocyte CEM cells placed in the upper chamber of a transwell chamber were assayed for chemotaxis in response to serial dilution of supernatant from DCs transduced with AdMDC, AdNull, or PBS alone (naive control) in the lower chamber. (b) EL4 cells. The study was similar to that in a, but the murine T lymphocyte EL4 cell line was used. (c) Suppression of AdMDC-mediated chemotaxis. CEM and EL4 cells were assayed for chemotaxis in response to 1:2 dilution of supernatant from AdMDC-modified DCs. Where indicated, anti–MDC neutralizing Ab or mouse IgG control Ab was added to the supernatant in the lower chambers at 10 μg/ml at the initiation of assay or cells were placed to the upper chamber in the presence of recombinant (r) MDC or TARC at 100 ng/ml. For all parts, the number of cells migrating to the lower chamber at 37°C for 4 hours was counted by FACS analysis. Migration index was calculated as the number of cells migrating to the conditioned media over the number of cells migrating to control medium. Results represent the mean ± SE (n = 3 per data point).

Article Snippet: To demonstrate the function of the MDC protein expressed by the AdMDC-modified DCs, human T lymphocyte CEM cells or murine T lymphocyte EL4 cells (no. CCL-119 and TIB-39, respectively; American Type Culture Collection, Manassas, Virginia, USA) were suspended in 100 μl of RPMI-1640 media containing 1 mg/ml BSA at 10 7 cells/ml and loaded to the upper wells of 5-μm pore, polycarbonate, transwell chambers in 24-well plates (Corning Costar Inc., Corning, New York, USA).

Techniques: Chemotaxis Assay, Genetically Modified, Serial Dilution, Transduction, Control, Modification, Recombinant, Migration

Journal: Journal of Biological Chemistry

Article Title: The Interferon-inducible p202a Protein Modulates NF-κB Activity by Inhibiting the Binding to DNA of p50/p65 Heterodimers and p65 Homodimers While Enhancing the Binding of p50 Homodimers

doi: 10.1074/jbc.m302105200

Figure Lengend Snippet: FIG. 2. p202a inhibited the binding to DNA of p50/p65 heterodimers but increased the binding of p50 ho- modimers in nuclear extracts from 293, HeLa, and EL4 cells. Aliquots of nuclear extracts from 293, HeLa, and EL4 cell cultures were incubated with 32P-la- beled RRBE oligodeoxynucleotide in 20 l of EMSA reaction mixtures in the absence of GST-202a (lanes 1, 5, and 9), or in the presence of 100 ng of GST-202 (lanes 2, 6, and 10), 200 ng of GST-202a (lanes 3, 7, and 11), or 400 ng of GST-202a (lanes 4, 8, and 12). After incubation, the reaction mixtures were analyzed by EMSA. The positions of the p50/p65 heterodimer and p50 homodimer bands as well as of the free probe are indicated. For further de- tails, see “Experimental Procedures.”

Article Snippet: EL4 (mouse T) cells (ATCC TIB181) were grown in suspension in RPMI medium supplemented with 10% fetal bovine serum, and 1% antibiotic and antimycotic agents (Invitrogen).

Techniques: Binding Assay, Incubation