Figure 4

Impedances of MCF7 cells transfected with SIN3A c.2830 C>T (A) MCF7 cells were transfected with indicated vectors together with pMACS 4.1 vector that expresses a truncated CD4 gene and pGFP expression vector for normalization for gene expressing cells. The untransfected MCF7 cells were also used as control cells. After transfection, the cells were cultured for 24 hrs and the transfected cells were separated with anti-CD4 antibody magnet beads. The separated cells were seeded at a density of 2.5 × 10⁵ cells/ml on a 16 well E-plate, and were incubated in the presence of 100 nM E2. Cell impedances were then continuously monitored at 15 min intervals for 24 hrs using the xCELLigence system. The data represent the means ± SE determined from 3 wells, and the figure shows representative data obtained from 2 independent experiments. (B) MCF7 cells were transfected with indicated vectors together with pMACS 4.1 vector that expresses a truncated CD4 gene and pGFP expression vector for normalization for gene expressing cells in the presence of control siRNA or siRNA for ESR1. The untransfected MCF7 cells were also used as control cells. After transfection, the cells were cultured for 24 hrs and the vector-transfected cells were separated with anti-CD4 antibody magnet beads. The separated cells were seeded at a density of 2.5 × 10⁵ cells/ml on a 16 well E-plate, and were incubated in the presence of 100 nM E2. Cell impedances were then continuously monitored at 15 min intervals for 24 hrs using the xCELLigence system. The data represent the means ± SE determined from 6 wells, and the figure shows representative data obtained from 3 independent experiments. (C) The normalized cell indexes were determined from data of Fig. 5A, and GFP fluorescence intensities were calculated using an IN Cell analyzer 2000 at 24 hrs from the start. The data represent the means ± SE determined from 3 wells. Statistical significances were determined by ANOVA with the Tukey–Kramer multiple comparison test (*P < 0.05). (D) The normalized cell indexes were determined from data of Fig. 5B as similar to Fig. 5C.