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3 protocols using lead 4 acetate

1

HEK-293 Cell Line Exposure to Lead Acetate

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The human embryonic kidney cell line, HEK-293, was obtained from American Type Culture Collection (ATCC, Manassas, VA). HEK-293 cells were cultured in Eagle's minimum essential media (EMEM, Gibco, Carlsbad, CA) supplemented with 10% Fetal Bovine Serum (FBS, SAFC BioScience), 5% penicillin streptomycin (Life Technologies, Carlsbad, CA), and 1% non-essential amino acids (Life Technologies, Carlsbad, CA). Cells (1×105) were plated in six-well plates, grown to 40-50% confluence and treated 24 hours post plating with lead (IV) acetate (Sigma; St. Louis, MO) at concentrations from 0-25 μg/dL. Seventy-two hours post treatment cells were 90-95% confluent and harvested. Genomic DNA was isolated using the Qiagen DNeasy Blood and Tissue Kit (Qiagen; Valencia, CA). Experiments were performed in four experimental replicates plated in triplicate for independent biological and technical replicates.
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2

Synthesis and Characterization of HP-β-CD Complexes

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HP-β-CD
with a molar substitution between 0.6 and 0.9 was kindly received
as a gift from Wacker Chemie AG (Germany). Sodium hypophosphite hydrate
(SHP, Sigma-Aldrich), BTCA (Sigma-Aldrich, 99%), PAHs [acenaphthene
(Ace, 99%), fluorene (Flu, 98%), fluoranthene (FluA, 98%), phenanthrene
(Phe, 98%), and pyrene (Pyr, 98%)], and heavy metals (zinc(II) acetate,
manganese(II) acetate, lead(IV) acetate, cadmium(II) acetate, copper(II)
acetate, and nickel(II) acetate) were all obtained from Sigma-Aldrich.
High-purity water was produced from a Millipore Milli-Q system (resistivity ≥
18 MΩ cm).
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3

Fabrication of Polycrystalline PbZrO3 Films

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Polycrystalline PbZrO3 films were prepared by chemical solution deposition using an inverted mixing order chemistry41 . The solution comprised zirconium butoxide (80 wt.% in butanol, Sigma Aldrich), lead (IV) acetate (Sigma Aldrich), methanol, and acetic acid. A 0.35 M solution was used with 35 mol% excess Pb to account for loss to the substrate and atmosphere during high-temperature annealing. The PZO film was coated on a 100 nm Pt/40 nm ZnO/300 nm SiO2/(001) Si substrate by spin casting at 4000 RPM for 30 s followed by a pyrolysis step at 350 °C for 1 min42 . After four coating and pyrolysis steps, an additional layer of a 0.1 M PbO solution was coated onto the surface to provide a PbO overpressure and the film was annealed at 700 °C for 10 min in a preheated box furnace43 . The final PbZrO3 film thickness was 300 nm. X-ray diffraction was performed using a PanAlytical Empyrean diffractometer in a Bragg–Brentano geometry with a GaliPIX detector and Cu Kα radiation, and scanning electron microscopy was performed using a FEI Quanta 650 with a concentric backscatter detector and an acceleration voltage of 15 kV44 .
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