Glibenclamide

Propofol (2,6-diisopropylphenol) and 2,4-diisopropylphenol were obtained from Sigma Aldrich. Glibenclamide was obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan), diazoxide from Abcam (Cambridge, MA, USA), and stromatoxin-1 from Alomone Labs Ltd. (Jerusalem, Israel).

Albendazole was purchased from Tokyo Chemical Industry Co., Ltd (Tokyo, Japan). Griseofulvin, dipyridamole, and glibenclamide were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). An identical batch was used by all companies.
The Japanese Pharmacopeia (JP) 2^nd fluid for dissolution testing (12.5 mM KH₂PO₄ and 12.5 mM Na₂HPO₄, pH 6.8, buffer capacity 10 mM/ ΔpH) was used for the solubility measurement.

Dimethyl sulfoxide (DMSO) was obtained from Junsei (Tokyo, Japan). N^G-nitro-L-arginine methyl ester (L-NAME), 4-aminopyridine (4-AP), tetraethylammonium (TEA), and glibenclamide were obtained from Wako Pure Chemical Industries (Osaka, Japan). 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) was obtained from Tokyo Chemical Industry (Tokyo, Japan). Krebs–Henseleit (KH) solution used in this study was composed of 118.0 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl₂, 1.2 mM MgSO₄, 1.2 mM KH₂PO₄, 11.1 mM D-glucose, and 25.0 mM NaHCO₃. All the reagents used in KH solution, BaCl₂, and urethane were obtained from Daejeong Chemical and Gold (Siheung-si, Republic of Korea). All other reagents were purchased from Sigma Aldrich (St. Louis, MA, USA).

ELC-org monolayers were incubated with HBSS containing 5 μM ¹⁴C-labeled glycyl-sarcosine (Glycyl-sarcosine, [1-14C]-, ARC) for 90 min at 37 °C. Accumulated glycyl-sarcosine in the monolayer was extracted by adding 70% methanol. Cell lysis solutions were measured by using a liquid scintillation counter (MicroBeta2; PerkinElmer). The PEPT1 activities were normalized with the protein content per well by using a Pierce BCA Protein Assay Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. As an inhibitor of PEPT1, 100 μM glibenclamide (FUJIFILM Wako Pure Chemical Industries) was used.

Antibody against Flag tag and CHX were purchased from Sigma-Aldrich (St Louis, MO, USA). Antibody against Myc tag was purchased from Life technologies (Rockville, MD, USA). Antibody against actin and IM-54 was purchased from Merck Millipore (Billerica, MA, USA). Antibodies against Abl (sc-23), phospho Tyr (PY99, sc-7020), caspase-1 p10 (sc-514), S100A8 (sc-8113), S100A9 (sc-8115) and caspase-1 inhibitor (z-YVAD-fmk) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against cleaved caspase-3 and cleaved PARP were purchased from Cell Signaling Technology (Danvers, MA, USA). Blocking antibody against mouse TNF and purified NA/LE hamster IgG1, λ1 isotype control was purchased from BD Biosciences (San Jose, CA, USA). Blocking antibody against mouse IL-1β was purchased from R&D systems. (Minneapolis, MN, USA). Imatinib mesylate was purchased from Novartis (Basel, Switzerland). Human TNF-α was purchased from Peprotech (Rocky Hill, NJ, USA). Q-VD-OPH was purchased from TONBO biosciences (San Diego, CA, USA). Glibenclamide was purchased from Wako (Osaka, Japan).