Mel 888

Human breast cancer cell lines MCF7 and MDA-MB-468, human melanoma cancer cell lines MEL-526 and MEL-888, human lung carcinoma cell line A549, human prostate cancer cell line PC3, and human colorectal adenocarcinoma cell line HT-29 were purchased from American Type Culture Collection (ATCC). MEL-526, MEL-888, and PC3 cells were cultured in RPMI-1640 (Hyclone). MCF7, MDA-MB-468, and A549 cells were cultured in Dulbecco’s Modified Eagle Medium (Hyclone). HT-29 was cultured in McCoy’s 5A Medium (Gibco). All cells were cultured with presence of 1X penicillin-streptomycin and supplemented with 10% heat-inactivated fetal bovine serum (Gibco) during normal growth conditions. Cells were cultured with antibiotic-free growth medium during drug treatment. Doxorubicin purchased from LC laboratories (#D-4000), anti-mouse PD-1 (#BE0033-2) and mouse IgG isotype control (#BE0086) from BioXCell. The PAK inhibitor PF-03758309 (PAKi) purchased from Selleckchem (#S7094). In nucleic acid depletion assay cells were treated with 10 µ/mL of DNase or 10 µ/mL of RNase (Invitrogen).

Human melanoma cancer cell lines MEL-526 and MEL-888, human breast cancer cell lines MCF7 and MDA-MB-468 were purchased from American Type Culture Collection (ATCC). MEL-526 and MEL-888 cells were cultured in RPMI-1640 (Hyclone) supplemented with 10% heat-inactivated Fetal Bovine Serum (HI-FBS) (Gibco). MCF7 and MDA-MB-468 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Hyclone). All cells were cultured with presence of 1X Penicillin-Streptomycin during normal growth conditions. During the drug treatments cells were cultured with antibiotic-free growth media. The PAK inhibitor PF-3758309 (PAKi) purchased from Selleckchem (Cat. #S7094). The drug was dissolved in dimethyl sulfoxide (DMSO). Cell lines were treated with 0 to 500 nM of PAKi for 48 h. In nucleic acid depletion assay cells were treated with 10 U/mL of DNase or 10 U/mL of RNase (Invitrogen).