Xcelligence real time cell analysis device

MCF7 and 4T1 cells were seeded at a starting density of 9 × 10³/well and 5 × 10³/well, respectively, in an electronic microtiter plate (E-Plate^®) (ACEA Biosciences, San Diego, CA, USA). The cells were cultured for 24 h before they were treated with 0, 5, or 10 µM DEA for 24 h, during which the impedance was measured every 5 min. The xCELLigence Real-Time Cell Analysis (RTCA) device (ACEA Biosciences, San Diego, CA, USA) was used according to the manufacturer’s protocol. The instrument was placed in a humidified incubator at 37 °C and 5% CO₂. These experiments were repeated twice running in three parallels.

To monitor the effects of HO-5114 on the growth of MCF7 and MDA-MB-231 cells, we used the xCELLigence system that allows for the real-time, quantitative analysis of adherent cells. The measurement method is based on the use of electronic microtiter plates (E-Plate^®), in the xCELLigence Real-Time Cell Analysis (RTCA) device (ACEA Biosciences, San Diego, CA, USA); both were used according to the manufacturer’s protocol. The instrument was placed in a humidified incubator at 37 °C and 5% CO₂. Cells were seeded at the starting density of 1 × 10³/well and were cultured for 24 h. Then, the cells were exposed to 75, 100, and 250 nM HO-5114 for seven days in the E-Plate^®, during which the impedance was measured each hour.

Cells were seeded at a density of 1 × 10³/well and cultured for 24 h before treatment with 2 µM olaparib, 25 µM oxaliplatin and 1 µM LY294002, alone and in combination, in an electronic microtiter plate (E-Plate^®) (ACEA Biosciences, San Diego, CA, United States). The treatment lasted 7 days. The impedance was measured every hour, using gold electrodes at the bottom of the wells. The xCELLigence Real-Time Cell Analysis (RTCA) device (ACEA Biosciences, San Diego, CA, United States) was used according to the manufacturer’s protocol. The instrument was placed in a humidified incubator at 37 °C and 5% CO₂.