Mda mb 231

All cell lines were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). MDA-MB-231, A549, and MIAPaCa-2 cells were cultured in high glucose Modified Eagle Medium (DMEM, Genesee Scientific, San Diego, CA, USA). NCI-H1299 cells were cultured in RPMI 1640 (Genesee Scientific, San Diego, CA, USA). All media were supplemented with 10% heat-inactivated fetal bovine serum (Genesee Scientific, San Diego, CA, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Genesee Scientific, San Diego, CA, USA). The cultures were incubated at 37 °C in 5% CO₂/95% humidified air. In all cases, treatment with experimental agents was done in basal medium supplemented with 5% heat-inactivated fetal bovine serum. Antibodies specific to B-Raf (Cat. #14814), Phospho-B-Raf (Ser445) (Cat. #2696), c-Raf (Cat. #53745), Phospho-c-Raf (Ser338) (Cat. #9427), MEK1/2 (Cat. #8727), Phospho-MEK1/2 (Ser217/221) (Cat. #9154), p44/42 MAPK (Erk1/2) (Cat. #4695), RSK1/RSK2/RSK3 (Cat. #9355), Phospho-p90RSK (Ser380) (Cat. #11989), GAPDH (HRP Conjugate) (Cat. #8884), α-Actinin (HRP Conjugate) (Cat. #12413), and anti-rabbit IgG, HRP-linked Antibody (Cat. #7074) were purchased from Cell Signaling Technology (Danvers, MA, USA).

All cells were obtained from American Type Culture Collection (ATCC). HEK293T, MDA-MB-231, MCF7, Hs 578T and their derived cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Genesee Scientific, 25–500). T-47D, MDA-MB-453, MDA-MB-468, BT-549, HCC70 and their derived cell lines were maintained in RPMI 1640 medium (Corning, 10-040-CV). 10%fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin were supplemented in the medium. GSK3326595 (HY-101563), MRT68921 dihydrochloride (HY-100006A), and Chloroquine (HY-17589A) purchased from MedChemExpress.