Anchorage-independent growth of tumor cells was estimated by soft agar growth assays. First, 1 × 104 transfected HCC cells were suspended using DMEM with 0.7% agar. Cells were plated on the top of a layer of 1.4% medium agar. Dishes were marked and incubated at 37°C for 10 days. We counted and photographed viable colonies (≥ 0.1 mm).

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