Anti pdgfra

Imatinib (Cayman Chemical, Michigan, USA) and Crenolanib (Selleckman) were dissolved in DMSO to give a final concentration of 10 mM, and used at the previously established IC₃₀ concentration, that is of 25 μM and 1 μM respectively. The following primary antibodies were used: primary mouse monoclonal anti-PDGFRB, anti-PDGFRA, and anti-FLT3 (Santa Cruz, TX, USA; 1:300) or mouse monoclonal anti-ß-actin (Anti-Actin, Clone C4, Millipore, MA, USA; 1:5000). IgG-HRP Santa Cruz (1:5000) was used as secondary antibody. Different silencing-RNAs (siRNAs) were tested, and purchased from Qiagen (Qiagen, UK). Further experiments were performed by using a single specific siRNA for PDGFRB (SI00605738, the so-called “siPDGFRB” now-on) and the “AllStars Negative Control siRNA” (SI03650318, used as non-targeting control - the so-called “C-PDGFRB” now-on). siRNA oligonucleotides were re-suspended in the provided buffer at a final stock concentration of 20 mM, and employed at 50nM in each experiments. HiPerfect transfection reagent was employed for siRNA trasfection (Qiagen, UK), as previously described [16 (link)].