Cim plate wells

The xCELLigence RTCA system (ACEA Biosciences, Roche, Germany) was used to understand the effect of miR-216b-5p on the proliferation rate of WT and TR in real time. Cells were seeded at 2 × 10⁴ cells/well containing 10 ug/mL trastuzumab in E-plate 16 (ACEA Biosciences). The cells were monitored every 30 min.
The migration and invasion assays were performed as previously described (Noyan et al., 2021 (link)). Briefly, cell invasion analysis was performed with xCELLigence real-time cell analyzer. The miR-216b-5p mimic transfected cells were seeded with serum-free medium into the upper chamber of the ACEA Biosciences Inc. CIM-plate wells (Cat. no: 2801038), which was covered with a microporous membrane of matrigel separating the upper and lower chamber. The lower chamber was filled with chemoattractant that is culture medium supplemented with 10% FBS. Negative control (NC) wells contained serum free medium as the control of the experiment. Both cell invasion and motility were monitored for 24 h with xCELLigence realtime cell analyzer, using CIM-plate and measuring impedance-based signals. The same experimental set up was used without matrigel for motility experiments.

Cell invasion analysis was performed with xCELLigence real-time cell analyser. At 24h post-transfection with miRNA mimic, 4x10⁴ BT-474 and SK-BR-3 cells in 200 uL of serum-free medium were seeded into the upper chamber of the ACEA Biosciences Inc. CIM-plate wells (Cat. No: 2801038), fitted with a microporous membrane of matrigel separating the upper and lower chamber. The lower chamber was filled with culture medium supplemented with 10% FBS as a chemoattractant. Cell invasion was monitored for 24 hours with xCELLigence real-time cell analyser, using CIM-plate and measuring impedance-based signals.