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. Author manuscript; available in PMC: 2017 Feb 1.
Published in final edited form as: Int J Chronic Dis Ther. 2016 Jun 23;2(3):31–38.

Figure 4.

Figure 4

Ubc9 knockdown suppresses the cell growth of ovarian cancer cell line UWB1.289 studied using phase contrast microscope (A) and with (B) Electric Cell-substrate Impedance Sensing (ECIS). The same starting cell density of UWB1.289 cells (shown in graphs) was seeded into the ECIS cultureware 8W10E+ array (Applied BioPhysics) and transfected with Ubc9 or control siRNA (100 nM) (Qiagen). The temperature of the cell suspension was prewarmed to the incubator temperature before the array was connected into the instrument electrode. After Setting up the software and the time course measurement for cell growth was conducted as described by the manufacturer. (C) Western blot analysis showing Ubc9 knockdown in ovarian cancer cell UWB1.289. Ubc9 or control siRNA (Qiagen) were transfected into UWB1.289 cells (1×106) using HiPerFect Transfection Reagent (Qiagen) after cells were seeded into the 10cm of Petri dishes 24 hours. Forty-eight hours later, the cells were collected and lysed in RIPA buffer. The cell lysates were probed with primary antibodies either Ubc9 (Santa Cruz N15, 1/100) or β Actin (Santa Cruz C4, 1/2000). The proteins were visualized using HRP labeled secondary antibody to goat (Ubc9) or mouse (β Actin) and developing solution (GE Healthcare) through Image Reader LAS-3000 (FUJIFILM).