Journal: Mutagenesis

Article Title: Mutagenicity of mercury chloride and mechanisms of cellular defence: the role of metal-binding proteins.

doi: 10.1093/mutage/15.6.525

Figure Lengend Snippet: Fig. 2. Intracellular GSH and MT content of 3T3 cells incubated for 6 h with Hg2. Results are presented relative to the control (100%) SD. The Fig. 4. Intracellular MT content of Hg2-treated NIH 3T3 cells (incubation absolute values of the controls are 1.2 mmol GSH/g protein and 47 mg time: 6 h) and changes resulting from an additional treatment with 5 µM MT/g protein. For each parameter two independent experiments were BSO. Bars represent the mean SD of two experiments, each performed in performed in triplicate. *Significantly different from the untreated control triplicate with the different combinations. *Significantly different from the (P 0.05). untreated control group (P 0.05); (a) significant difference between the Hg-treated and the HgBSO-treated cells (P 0.05).

Article Snippet: The Cd content of the sampleCell culture allows direct calculation of its MT content (Onosaka and Cherian, 1982),NIH 3T3 cells (ATCC no. CRL 1658) were subcultured at 37°C in a 5% which was related to the total protein content measured.CO2 atmosphere in DMEM supplemented with 10% fetal calf serum, 2 mM L-glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin.

Techniques: Incubation, Control

Journal:

Article Title: Loss of Protooncogene c-Myc Function Impedes G 1 Phase Progression Both before and after the Restriction Point

doi: 10.1091/mbc.E02-10-0649

Figure Lengend Snippet: Single BrdU labeling (A and B) and double CldU/IdU labeling (C) of asynchronously cycling cells. (A) Pulse labeling with BrdU. Cells were pulse labeled for the indicated times, harvested immediately afterward, and processed as described in MATERIALS AND METHODS. (B) Continuous labeling with BrdU. BrdU was added at t = 0 and cells were harvested at the indicated time points. The labeling indices (means ± SD, n = 11 fields/time point) denote BrdU-positive cells as percentage of total cells. A minimum of 687 c-myc+/+ and 676 c-myc−/− cells/time point (A) and 645 c-myc+/+ and 634 c-myc−/− cells/time point (B) were scored. (C) Double labeling with CldU and IdU. Cells were pulse labeled with CldU for 30 min at t = 0, and culture was continued in the absence of CldU as described in MATERIALS AND METHODS. At the indicated time points, the second pulse was delivered with IdU for 30 min and the cells were harvested immediately afterward. The labeling indices (means ± SD, n = 3–4 [c-myc+/+] and 6–10 [c-myc−/−] fields/time point) denote CldU/IdU double-labeled cells as percentage of total CldU-positive cells. A minimum of 57 c-myc+/+ and 44 c-myc−/− CdlU-positive cells/time point were scored. The BrdU labeling indices in this figure were obtained using in situ immunohistochemistry methods.

Article Snippet: IdU was detected with the mouse monoclonal anti-BrdU antibody (clone B44; catalog no. 347 580; BD Biosciences, San Jose, CA; Gratzner, 1982 ) diluted 1:10 in PBSA-T (2.5 μg/ml), followed by Alexa 488-conjugated goat anti-mouse secondary antibody (catalog no. A-11029; Molecular Probes, Eugene, OR) diluted 1:600 (3.0 μg/ml).

Techniques: Labeling, In Situ, Immunohistochemistry