The cells were exposed to Pb2+ either alone or in combination with CS-OREC composites for designated time to evaluate the efficacy of the composites on Pb2+-induced toxicity.
Cells were cultured in 6-well plates and infected with adenovirus-LETM1, adenovirus-GFP for designated time points.
Fibroblasts were trypsined and plated on 6-well plastic dishes pre-coated with 1 µg/ml fibronectin and incubated at 37°C in 5% CO2 incubator for designated time periods.
After being treated with I-Trp (5 μM) for designated time periods, cells were collected and resuspended in cell lysis buffer (BioVision).
Ovarian cancer cells in 24-well culture plates were treated with or without G-1 at the indicated concentrations for designated time points as noted in the figure legends.
For salt or ABA treatment, the shoots were transferred to beakers containing fresh distilled water added with either 200 mM NaCl or 100 μM ABA, kept in the same growth chamber for designated time (0, 5, 24, 48 and 72 h for salt, 0, 1, 2, 3, 4 and 5 d for ABA).
After cultivation without food for designated time (see below), animals were collected with NG buffer kept at 20 °C (except for Fig. S1 in Supporting Information) or at 23 °C (only for Fig. S1 in Supporting Information), and approximately 40 300 animals were placed at the center of the TTX plate.
Pellets were resuspended in 20 μl of TBP buffer (5 mM TBP, 8 M urea, 2% CHAPS, 20 mM Tris-HCl, pH 8.0) for 1 h at room temperature and then incubated in the dark with alkylating antitumor drugs for designated time.
Samples were incubated at 37°C for designated time periods, and the reaction was stopped by the addition of SDS PAGE loading dye, followed by heating at 100°C for 10 min. CXCR2 was detected by western blotting and mouse anti-His antibodies.
For designated time periods, the two kinds of enzymes retained more than 85% of their initial activity at 40°C for 30 min. However, at 60°C, PEL168E completely lost its activity after 20 min pre-incubation, whereas PEL168P kept 26% of its initial activity with the same treatment.
