Rpmi 1640 culture media

4ME was isolated and supplied by the Department of Natural Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences Brno, Czech Republic. The identification of substance was carried out using HRMS, ¹H and ¹³C NMR analysis; and the purity exceeded 95% according to the HPLC analysis [6 (link)]. The compound is poorly soluble in water; therefore, a fresh 10 mM stock solution in dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA) was prepared every time 1 day prior to experiments and stored at –20°C. This solution was further diluted in the culture media to the desired final concentrations. RPMI 1640 culture media, phosphate buffered saline (PBS), and antibiotics (penicillin and streptomycin) were purchased from Lonza (Verviers, Belgium). Foetal bovine serum (FBS) was purchased from PAA Laboratories (Pasching, Austria). Rabbit polyclonal antibodies against p38 MAPK [pT180/Y182] (9215S) and p38 MAPK (9212) were purchased from Cell Signaling Technologies (Beverly, MA, USA). Mouse monoclonal antibodies against pRb (554136) were purchased from BD Biosciences (Franklin Lakes, NJ, USA). Rabbit polyclonal antibody against phospho-Rb [Ser 780] (9307) was purchased from Cell Signaling Technologies (Beverly, MA, USA). PE-conjugated CD11b antibody was obtained from Beckman Coulter (Brea, CA, USA). ATRA and all other reagents were from Sigma-Aldrich.