Anti gpr55

DMEM/F12, DMEM, IMDM, L-glutamine, HCl, Earle’s salts solution, Hank’s salts solution, Versene’s solution, antibiotic/antimycotic mixture (penicillin, streptomycin, amphotericin B), DMEM, trypsin, and fetal bovine serum were from Servicebio, Hubei, China.
Cell lines MDA-MB-231 (HTB-26), MCF-10A (CRL-10317), MCF-7 (HTB-22), BT-474 (HTB-20), BT-20 (HTB-19), HL-60 (CCL-240), LN-229 (CRL-2611), and SK-BR-3 (HTB-30) were purchased from ATCC, Manassas, VA, USA.
Antibodies anti-b-actin, anti-CB1, anti-CB2, and anti-GPR55 were from Abcam, Cambridge, UK. Anti-mouse IgG antibody was from Jackson ImmunoResearch, Cambridge, UK.
SR 144028, PSB CB5, ML-184, ML-193, and LPI were from Tocris Bioscience, Bristol, UK. DMSO, resazurin, NAD⁺, DL-lactate, diaphorase, iodonitrotetrazolium chloride (INT), D-glucose, glycylglycine, acetic acid, MgSO_4, EGTA, dithiothreitol, DMSO, Triton X-100, acrylamide, bis-acrylamide, Triton X-100, SDS, nitro blue tetrazolium, Tris, EDTA, agarose, bicinchoninic acid, D-glucose, bovine serum albumin, anti-rabbit IgG antibody, and 5-Bromo-4-chloro-3-indolyl phosphate-toluidine were from Sigma-Aldrich, St. Louis, MO, USA. The purity of all used reagents was 95% or more.
rhEGF was from SciStoreLab, Skolkovo, Russia.

For Western blot analysis, the total protein samples (80 μg/well) were loaded and separated by SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), then transferred to a PVDF membrane. The membrane was blocked in blocking buffer (5% w/v BSA in Tris-buffered saline (25-mM Tris, pH 7.4, 150-mM NaCl, and 0.05% Tween 20) (TBS-T) for 1 h at room temperature, then incubated in appropriate primary antibodies for 1 h at room temperature or overnight at 4 °C. The primary antibodies used were: anti-ABCC1 derived in rabbit (1:1000, EPR4658(2); Abcam), anti-ABCC4 derived in rat (1:100, M₄I-10; Abcam), anti-ABCB1 derived in mouse (1:200; Thermo Fisher), anti-ABCG2 derived in mouse (1:50, Bxp1; Santa Cruz, Heidelberg, Germany), or anti-GPR55 (rabbit) (1:100; Abcam). The secondary antibodies used were anti-rabbit HRP (1:3000; Cell Signaling Technology, Leiden, The Netherlands), anti-rat HRP (1:5000; Merck), and anti-mouse HRP (1:4000; Cell Signaling Technology). Blots were visualized using a SuperSignal West Chemiluminescent kit (Pierce, Thermo Fisher) and the Li-Cor C-Digit blot scanner, and the images were analyzed using the Image Studio Lite software imaging system (Li-Cor, Cambridge, UK). Afterwards, blots were re-probed using anti-α-tubulin (mouse) (1:1000; Merck) to check for equal sample loading. Western blot analysis was repeated at least 3 times for each experiment.