Sodium chloride (nacl)

The cells were lysed with cell lysis buffer (pH 8.0) containing 50 mM Tris-HCl (Sigma-Aldrich), 150 mM NaCl (Guangzhou Chemical Reagent Factory, Guangzhou, China), 5 mM EDTA (Guangzhou Chemical Reagent Factory), 1% NP40 (Sigma-Aldrich), 0.05% phenylmethanesulfonyl fluoride (Sigma-Aldrich), 2 μg/ml aprotinin (Sigma-Aldrich) and 2 μg/ml leupeptin (Sigma-Aldrich). The protein levels were determined by western blotting, as described previously (4 (link)).

All solvents used were chromatographic grade. 1-Pentanol, employed as an internal standard, and a mixture of n-alkanes (C7-C30) used for the retention index (RI) analyses were purchased from Sigma-Aldrich (Steinheim, Germany). NaCl of analytical grade was purchased from Guangzhou Chemical Reagent Factory (Guangzhou, China), and water was purified using a Milli-Q system (Millipore, Billerica, Massachusetts).

Chemical reagents used for SG induction included sodium arsenite (AS), clotrimazole (CZ), H₂O₂, sorbitol (all from Sigma-Aldrich, Shanghai, China), and NaCl (Guangzhou Chemicalreagent Co., Ltd, Guangzhou, China). SGs were induced by treatment with AS (500 μM for 1 h, unless specifically indicated in the figure legends), CZ (20 μM for 1 h), H₂O₂ (1 mM for 1 h), NaCl (0.2 M for 30 min), or sorbitol (0.4 M for 30 min), or recovered from stress for 1 h. Cells were scored for SGs by manual counting using fluorescence microscopy with G3BP and FXR1 or eIF4G as SG markers; only cells with granules co-stained for these markers were considered SGs, and a minimum of three granules per cell was required for a positive score. In the experiments with Flag-tagged UBAP2L deletants or mutants, only Flag-positive cells were considered, and the number of granules in an individual cell was counted. At least 100 cells from at least five fields were analyzed.

Minimum essential medium (MEM), fetal bovine serum (FBS), penicillin/streptomycin, Hanks Balanced Salt Solution, sodium pyruvate and trypsin were purchased from Gibco (Carlsbad, CA, United States). 5-Alpha-dihydrotestosterone (5α-DHT), collagenase IA, pyridoxal 5-phosphate, O-(carboxymethyl) hydroxylamine hemihydrochloride chloride (AOAA), DL-propargylglycine (PAG), sodium hydrosulfide hydrate (NaHS × H₂O), sodium lactate, L-cysteine (L-Cys), bumetanide, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), glibenclamide (Glib), Iberiotoxin (IbTx) and tetraethylammonium chloride (TEA) were purchased from Sigma-Aldrich (St. Louis, MO, United States). HC067047 was purchased from Tocris (Bristol, United Kingdom). NaCl, KCl, MgSO₄, FeCl₃, HCl, BaCl₂, NaHCO₃, KH₂PO₄, CaCl₂, glucose, mannitol, lactic acid, normal saline, N,N-dimethyl-p-phenylenediamine sulfate, trichloroacetic acid, and zinc acetate were purchased from Guangzhou Chemical Pharmaceutical Factory (Guangzhou, China). Universal two-step detection kit (PV-9000) and DAB detection kit were purchased from ZSBIO (Beijing, China). Fluo-3 AM was purchased from Molecular Probes (Eugene, OR, United States). PBFI AM was purchased from Cayman Chemical (Ann Arbor, MI, United States).