L02 and Hep3B cells were plated in 96-well flat-bottom tissue culture plates (Falcon) at a density of 5,000 cells per well. For acetaldehyde experiments, cells underwent acetaldehyde treatment as previously described, following a 24-hour plating period. At the 36-hour acetaldehyde treatment time point, cells were transfected with the miRNA inhibitors. Cellular proliferation was analyzed using an MTS proliferation assay (Promega) in accordance with the manufacturer’s protocol, beginning 24 hours after the last acetaldehyde treatment.
For doxorubicin sensitivity experiments, cells were transfected with the miRNA inhibitors after a 24-hour plating period. They were subsequently exposed to one of several doses of doxorubicin ranging from 0–4 μg/mL [20 (link), 21 (link)]. After a 48-hour incubation period, cellular proliferation was analyzed using an MTS proliferation assay (Promega). All assays were performed in triplicate wells and experiments were individually performed at least twice.
For doxorubicin sensitivity experiments, cells were transfected with the miRNA inhibitors after a 24-hour plating period. They were subsequently exposed to one of several doses of doxorubicin ranging from 0–4 μg/mL [20 (link), 21 (link)]. After a 48-hour incubation period, cellular proliferation was analyzed using an MTS proliferation assay (Promega). All assays were performed in triplicate wells and experiments were individually performed at least twice.
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