Ropivacaine

Bupivacaine, lidocaine and ropivacaine were obtained from Sigma-Aldrich (St. Louis, MO, USA). LevoBupivacaine was purchased from Abbott Laboratories Services Corp, Taiwan Branch (Taiwan). All other reagents used were of reagent grade.

The human MDA-MB-231 TNBC cells, human breast cancer BT-474 cells, and human normal gingival epithelial SG cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Corning, Corning, NY, USA), containing 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (Invitrogen, Waltham, MA, USA). The lidocaine, bupivacaine, levobupivacaine, ropivacaine, acridine orange, and doxorubicin were purchased from Sigma–Aldrich (St. Louis, MO, USA). The autophagy inhibitor 3-methyadenine (3-MA) was purchased from InvivoGen (San Diego, CA, USA).

Human mammary epithelial cell line MCF-10A, Her2+ SK-BR-3 cell lines, luminal BT474 and MCF-7 cell lines, and TNBC MDA-MB-231 and MDA-MB-468 cells lines were obtained in American Type Tissue Culture Collection. Cells were incubated at a condition of 37°C with 5% CO₂ in DMEM (Gibco, USA) with 100 units/mL penicillin (Gibco, USA), 0.1 mg/mL streptomycin (Gibco, USA), and fetal bovine serum (10%, Gibco, USA). Ropivacaine (Sigma, USA) was dissolved by DMSO and diluted in PBS. The miR-27b-3p mimic and inhibitor were purchased (GenePharma, China). The pcDNA3.1-YAP overexpression vectors and YAP siRNA were obtained using full-length synthesis (GenePharma, China). Liposome 3000 (Invitrogen, USA) was utilized for the transfection.

Human melanoma cell lines A375 and A431 (Cell Lines Service, Germany) were cultured in RPMI 1640 medium (Invitrogen, US) supplemented with 2 mM glutamine and 10% heat-inactivated fetal bovine serum (Gibco, US). Ropivacaine and bupivacaine (Sigma, US) were dissolved in water and lidocaine was reconstituted in Hanks Balanced Salt Solution. Veratridine (R&D Systems, US), vemurafenib (LC Laboratories, US), calpeptin (Sigma, US) and dacarbazine (Selleckchem, US) were reconstituted in dimethyl sulfoxide (DMSO). Tetrodotoxin (Sigma, US) was dissolved in citrate buffer.

Methanol (HPLC grade) was purchased from Honeywell Reagents (Seelze, Germany). Acetic acid (100% glacial), hydrochloric acid (37%), and ammonia solution (32%) were purchased from Merck (Darmstadt, Germany). n-octanol (99.5%), ammonium acetate (98.1%), ascorbic acid (99.5%), ⍺-tocopherol formulation (T 4389; approximately 400 mg ⍺-tocopherol per Gram), 2,3,5-triphenyl tetrazolium chloride (TPTZ, 98%), iron (III) chloride hexahydrate (98%), potassium persulfate (99%), 2,2-diphenyl-1-picrylhydrazyl (DPPH; 95%), lidocaine (98%) and ropivacaine (100%) were obtained from Sigma-Aldrich (Steinheim, Germany). 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS, 99%) was purchased from Biochemics, Fluka (Steinheim, Germany). Bupivacaine (99%) was purchased from Biosynth Crbosynth (Compton, United Kingdom). lidocaine Chlorhydrate (400 mg/20 mL, LOT 18T1397) was obtained from Braun. ropivacaine Hydrochloride (75 mg/10 mL, LOT 12PCA07) and Levobupivacaine hydrochloride (50 mg/10 mL, LOT 18T1397) were obtained from Fresenius Kabi. A Milli-Q water purification system (Millipore, Bedford, Massachusetts, United States) was used to obtain ultrapure water (18 MΩ cm⁻¹). Disposable plastic cuvettes were purchased from Brand (Wertheim, Germany).

Ropivacaine, 3-hydroxy Ropivacaine, and diazepam-d5 (internal standard), all with purities > 98% (Figure 1), were purchased from Sigma-Aldrich Co. LLC (St. Louis, MO, USA). Acetonitrile and methanol (HPLC grade) were purchased from Merck (Darmstadt, Germany). Ultrapure water was prepared by the Millipore Milli-Q purification system (Bedford, MA, USA).