Ma5 14336

BCL1 cells, grown in culture plates, were treated with or without Pt(S-pr-thiosal)2 (0.05 mg/mL) for 12 h. The treated cells were fixed and permeabilized with permeabilization buffer (BD Bioscience) and incubated with antibodies specific for Bcl-2 (11-6992-42, Thermo Fisher Scientific, Cambridge, UK), Noxa (ab13654, Abcam, Cambridge, UK), STAT3 (IC1799G, Novus Biologicals, San Diego, CA, USA), p16 (ab211542, Abcam, Cambridge, UK), p21 (ab188224, Abcam) and p27 (ab215434, Abcam), cyclin D3 (ab28283, Abcam Cambridge, UK), cyclin E (MA5-14336, Thermo fisher scientifics, Waltham, MA, USA), Ki-67 (151212, BioLegend, San Diego, CA, USA), Noxa (ab13654, Abcam) and Mcl-1 (ab32087, Abcam). For staining p16, p21, and p27 cells were additionally incubated with secondary goat anti-mouse IgG FITC (ab6785, Abcam) or donkey anti-rabbit IgG (ab150073, Abcam). Flow cytometry was performed on FACSCalibur flow cytometer (BD Biosciences). The data were analyzed using FlowJo (Tree Star).

4T1 cells, grown in culture plates, were treated with either DE-EDCP or cisplatin (31.25 μM) for 24 hours. Next, the treated cells were fixed and permeabilized with permeabilization buffer (BD Bioscience) and incubated with antibodies specific for STAT3 (IC1799G, Novus Biologicals, San Diego, USA), Ki-67 (11-5698-82, eBioscience, San Diego, USA), cyclin D3 (ab28283, Abcam Cambridge, United Kingdom), cyclin E (MA5-14336, Thermo fisher scientifics, USA), p16 (ab211542, Abcam Cambridge, United Kingdom), p21 (ab188224, Abcam Cambridge, United Kingdom) and p27 (ab215434, Abcam Cambridge, United Kingdom). For staining cyclin D3, cyclin E, p16, p21 and p27 cells were additionally incubated with secondary goat anti-mouse IgG FITC (ab6785 Abcam Cambridge, United Kingdom) or donkey anti-rabbit IgG (ab150073 Abcam Cambridge, United Kingdom). Flow cytometry was conducted on FACSCalibur flow cytometer (BD Biosciences, San Jose, USA) and the data were analyzed using FlowJo (Tree Star).