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. 2018 Jun 26;8:9706. doi: 10.1038/s41598-018-28010-y

Figure 2.

Figure 2

Sodium orthovanadate (SOV) reduces ATPase activity in sorafenib-resistant HCC cells. (a) HepG2-SR and Huh7-SR cells were incubated for 30 min with SOV at various concentrations. Cells were harvested and lysed, and the amounts of phosphate release were measured. (b,c) Cells were incubated with vehicle or SOV at 5 μM for 6, 12 or 24 h, and then stained with K+ fluorescent dye PBFI-AM. (b) Fluorescence intensity was measured. (c) Representative images were taken from PBFI-AM-stained cells incubated with vehicle or SOV (5 μM) for 24 h. (d,e) Cells were incubated with 5 μM of SOV for 24 h, and then subjected to qRT-PCR for measuring the expression of Na+/K+-ATPase α3 subunit mRNA (d) and immunoblotting for its protein expression (e). The density of each immunoblotting band was normalized to β-actin. “N.S.” indicates no significance. “*” (P < 0.05) and “**” (P < 0.001) vs. vehicle-treated cells.