Ethyl 3 aminobenzoate methanesulfonate tricaine

Articaine hydrochloride (ATC) was donated by DFL Ind. Com. S.A. (Rio de Janeiro, Brazil). Cetyl palmitate (CP, cat 501427) and Dhaykol 6040 LW (DK, cat SO1253) (caprylic/capric acid triglycerides) were purchased from Dhaymers Química Fina (São Paulo, Brazil). Pluronic F68 (P68, cat K4894), Tween 80 (T80, cat P1754), copaiba oil (CO, cat W521809), methyl cellulose (cat M0512) and ethyl 3-aminobenzoate methanesulfonate (Tricaine, cat E10521) were supplied by Sigma–Aldrich (St. Louis, MO, USA). Avocado butter and olive oil were purchased from Engenharia das Essências Ltda (São Paulo, Brazil). Liss rhodamine-PE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl)) was from Avanti Polar Lipids, Inc. (Alabaster, AL, USA, cat 810150). Low melting point agarose was obtained from Cleaver Scientific Ltd (Warwickshire, UK, cat CSL-LMA5). E3 medium (5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl₂; 0.3 mM MgCl₂, calibrated to pH 7 with NaHCO₃) was prepared at the Fish Immunology lab, Andres Bello University. Deionized water (18 MΩ) was obtained with an Elga USF Maxima ultra-pure water purifier.

Zebrafish lines in this study (Table S2) were handled in compliance with local animal welfare regulations, as overseen by the Animal Welfare Body of Leiden University (License number: 10612) and maintained according to standard protocols (zfin.org). Generation of the dram1 mutant was approved by the Animal Experimentation Committee of Leiden University (UDEC) under protocol 14198. All experiments were done on embryos or larvae up to 5 days post-fertilization, which have not yet reached the free-feeding stage. Embryos/larvae were kept in egg water (60 µg/ml Instant Ocean sea salts) containing 0.003% 1-phenyl-2-thiourea (PTU, Sigma-Aldrich) at 28.5 °C and treated with 0.02% ethyl 3-aminobenzoate methanesulfonate (Tricaine, Sigma-Aldrich) for anesthesia before bacterial injections, imaging and fixation.