Brdu 7aad flow kit

End-point cell viability assays were performed by using either Cell Titer-Glo cell viability assay kit (Promega) or WST-1 reagent (Roche, Penzberg, Germany) as manufacturer’s protocol and were measured using Synergy HT microplate reader (BioTek, Vermont, USA). Real-time cell viability, migration and invasion assays were done on xCELLigence RTCA-DP (ACEA Biosciences Inc., CA, USA) according to manufacturer’s protocol. All RTCA measurements were normalized to transfection time, and statistical significance of the results was tested by paired two-tailed student t-test. Cell cycle analysis was performed using BrdU/7AAD flow kit (BD Biosciences, USA) as previously described28 (link). For wound healing assay, 2.5 × 10⁵ cells per well were seeded in 24-well plates, and after 48 hours of incubation following the transfections and scratch formation, images were taken with 5x magnification using Nikon Eclipse inverted microscope (Nikon, Japan). All graphics and statistical analysis were carried out in GraphPad software (GraphPad software Inc., La Jolla, CA, USA).

EdU staining was done using Click-iT™ EdU Cell Proliferation Imaging Kit (ThermoFisher Scientific) according to manufacturer’s instructions. Annexin V/DAPI staining was done as previously described^{68 (link),69 (link)}. For assessing the effects of SOC on G1 arrest, T47D cells treated with 5 µM of SOC for 24 hours were fixed in ethanol for 30 min on ice and stained with DAPI. For analyzing cell cycle distribution upon SOC treatment at early time points, BrdU/7AAD Flow kit (BD Biosciences) was utilized based on manufacturer’s instructions.