Sodium azide bioxtra

Standard chemicals, 2-thenoyltrifluoroacetone (TTFA; #T27006), rotenone (#R8875), sodium azide BioXtra (#S8032), antimycin A (#A8674), dimethyl malonate (#04011), d-α tocopherol succinate (#T3126), 2,6-dichloroindophenol sodium salt hydrate (DCPIP; #D1878), phenazine methosulphate (PMS; #P9625), MTT (#M5655), acetyl CoA (#A2181), 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB; #D218200), bovine serum albumin (#A6003), doxycycline hyclate (#D9891), 3-Nitropropionic acid (#N5636) and oxaloacetic acid (#O4126) were purchased from Sigma-Aldrich. Carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone (FCCP; #BML-CM120-0010) was purchased from Enzo Life Sciences. Venetoclax (#2253-1) and IACS-010759 (#B2231-1) were purchased from Biovision Inc. Potassium phosphate monobasic (KH₂PO₄; #BP362-500) and ethylenediamine tetra acetic acid (EDTA; #BP118-500) were purchased from Fisher Scientific. Decylubiquinone was purchased from Abcam (#ab145233). Oligomycin was purchased from Merck Millipore (#495455). Triton X-100 (0694) was purchased from Amresco. Polybrene (#TR-1003-G) was purchased from Millipore. Puromycin (#631306) was purchased from Clontech.

Randomly selected subgroups of animals of each experimental group (Paired, n = 15; Unpaired, n = 10; and Saline, n = 10) were anesthetized with sodium pentobarbital (15 mg/kg) (Dolethal 100 mL; Vetoquinol E.V.S.A., Madrid, Spain) 90 min after the preference test and perfused transcardially, first with heparin solution (0.006%) (Heparin sodium salt from porcine intestinal mucosa; Cat# H3393; Sigma-Aldrich, Madrid, Spain) and then with paraformaldehyde (4%) (Paraformaldehyde, powder, 95%; Cat# 158127; Sigma-Aldrich, Madrid, Spain). After perfusion, each brain and cerebellum were dissected and placed in a container with the same fixative at room temperature. Twenty-four hours later, we immersed the tissue in a 30% sucrose solution in phosphate buffered (0.10 M) with sodium azide (2%) (Sodium azide BioXtra; Cat# S8032; Sigma-Aldrich, Madrid, Spain) and preserved at 4°C until the brains sunk. Then, we sliced the brain tissue into 40 μm-sections and collected four series using a cryostat microtome (Microm HM560 Thermo Fisher Scientific, Barcelona, Spain), storing them at 20°C in a cryoprotectant solution. Then, we selected sagittal sections (comprising the whole vermis) of LVIII and lobule IX (LIX) of the cerebellum (Figure 2A) and coronal sections of VTA (Figure 3A), NAc (Figure 3A), and mPFC (Figure 3A) for subsequent immunofluorescence.