Ethylisopropyl amiloride

Two percent sodium alginate was prepared by dissolving sodium alginate (Sigma-Aldrich) in phosphate-buffered saline (PBS) and sterilizing by heating to 80°C in a water bath for 15 min (Leo et al., 1990 (link)). Anti-CD150-PE, anti-GRPC5C Alexa Fluor-405, and anti-Ki67-Alexa Fluor 405 antibodies for flow cytometry were purchased from R&D Systems. Anti-CD49f-PE and anti-CD90-VioBlue were purchased from Miltenyi Biotec. Transforming growth factor beta-1 (TGFβ-1) and fibroblast growth factor were obtained from PeproTech. Fms-like tyrosine kinase-3 (Flt-3), G-CSF, stem cell factor (SCF), and interferon alpha were purchased from ImmunoTools. All-trans retinoic acid (ATRA) was obtained from Sigma-Aldrich. Hoechst 33342, propidium iodide (PI), and pyronin Y (PY) were purchased from Sigma-Aldrich. The CountBright^TM Absolute Counting Beads was purchased from Thermo Fisher Scientific. MHY1485, AZD8055, rosiglitazone, KU-55933, LEE011, roscovitine (seliciclib, CYC202), glasdegib (PF-04449913), sodium butyrate, dasatinib, BIO, plerixafor (AMD3100), quizartinib, and sorafenib were purchased from Selleckchem. Adiponectin, triglitazone LE135, PD169316, harmine, nilotinib, ethylisopropyl amiloride, anisomycin, and curcumin were purchased from Sigma-Aldrich, while prostaglandin E2 was from BioVision/Cambridge Bioscience. Cytarabine and daunorubicin were purchased from Sigma-Aldrich.

Roswell Park Memorial Institute (RPMI) 1640 medium, bafilomycin A1 (an inhibitor of V-type H⁺-ATPase), ethyl isopropyl amiloride [EIPA; an inhibitor of Na⁺/H⁺ exchanger (NHE)], acridine orange (AO) and valinomycin were purchased from Sigma-Aldrich (St. Louis, MO, USA). N-(Ethoxycarbonyl methyl)-6-methoxy quinolinium (MQAE) and carboxy-seminaphthorhodafluor-1 (carboxy-SNARF-1) were obtained from Invitrogen (Carlsbad, CA, USA). Nigericin, tributyltin chloride and monensin were purchased from Wako Pure Chemical Industries (Osaka, Japan). The moderately differentiated human gastric adenocarcinoma cell line, MKN28, was obtained from ATCC (Manassas, VA, USA).

All cell lines used were obtained from ATCC (Manassas, VA) and have since been validated (Promega). DU145 (prostate carcinoma), H1993 (lung carcinoma), AU565, and HCC1806 (breast carcinoma) cells were maintained in RPMI 1640 media (Cellgro, Herndon, VA), MDA-MB-231 (breast carcinoma) cells were maintained in DMEM (Cellgro), and HEK293 cells in EMEM (Cellgro); each containing 10% fetal bovine serum (FBS) (Gemini, West Sacramento, CA). Cells were kept at 37°C with 5.0% CO₂. Monensin and brefeldin A were obtained from Biolegend (San Diego, CA), Y27637 from Stem Cell Technology (Vancouver, BC), C75, 2-hydroxymyristic acid, and 17-Octadecynoic Acid from Caymen Chemicals (Ann Arbor, MI), cycloheximide, recombinant EGF, geranylgeranyltransferase inhibitor, farnesyltransferase inhibitor, vinblastine, latrunculin, dansylcadaverine, ethylisopropyl amiloride, methyl-β-cyclodextrin, nystatin, and 2-bromopalmitate were obtained from Sigma-Aldrich (St. Louis, MO). Click-iT® reaction buffers, azido-homoalanine, azide-palmitate, and methionine-free RPMI, were obtained from Invitrogen (Carlsbad, CA). Recombinant HGF was obtained from EMD Millipore (Darmstadt, Germany). Streptavidin Sepharose High Performance beads were obtained from GE Healthcare (Pittsburgh, PA).