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Parental Huh-7 HCC cells and the sorted cells after Fluorescence Activated Cell Sorting (FACS, FACSAria III, BD) were maintained in complete growth medium (Dulbeccos modified Eagle medium, DMEM, Sigma) containing 10 heat-inactivated fetal bovine serum (FBS, Biowest), 2 mM L-glutamine (Sigma) and 0.1 mM non-essential amino acids (Sigma).Enhanced expression of STIM1 in cancer tissue has been associated with poor patient prognosis.Methods We performed qRT-PCR, intracellular Ca 2 monitoring, protein analyses, and real-time cell proliferation assays on EpCAM()CD133() subpopulation of tumor-initiating Huh-7 HCC cells expressing high levels of STIM1 andor Orai1.
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