Western blotting was conducted as described [23 (link)]. Polyclonal rabbit antibody for Fli-1, PKCδ, Phospho-PKCδ, PKCα, Phospho-PKCα were obtained from Abcam (Abcam, Cambridge, UK); ERK, phospho-ERK and GATA1 antibodies from Cell Signalling Technology (CST, Danvers, MA01923), GAPDH from Sigma (Sigma Aldrich, China) and β-actin from Proto-Technology (Protein-Tech, Bucuresti, Romania). The dilution of antibodies was used according to the manufacturer instructions. The inhibitors of PKC (GO6983), MEK (U0126), RAF (Sorafenib Tosylate), JNK (SB600125), Calcimycin, Rottlerin, and P38 (SB203580) were all obtained from Sellectchem (Sellectchem, Houston, USA). These compounds were dissolved in DMSO and added to the cells at indicated concentration, as described in the results section.
For immunoprecipitation, Extracts isolated from HEL and CB7 cells were immunoprecipitated using anti-Fli-1 antibody followed by western blot with anti-phospho Serine/Threonine antibody (Abcam), as previously described [23 (link)].
For immunoprecipitation, Extracts isolated from HEL and CB7 cells were immunoprecipitated using anti-Fli-1 antibody followed by western blot with anti-phospho Serine/Threonine antibody (Abcam), as previously described [23 (link)].
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