Absolute ethanol

Thiobarbituric acid (TBA), Trichloroacetic acid (TCA) and 3,5-Di-tert-4-butylhydroxytoluene (BHT) acquired from Sigma-Aldrich (Steinheim, Germany). These reagents were used for lipid peroxidation determination. Especially, BHT was used as an antioxidant agent in this procedure.
Absolute ethanol, acetone and n-hexane were obtained from Panreac Química (Barcelona, Spain), Scharlau (Barcelona, Spain) and Macron (Center Valley, PA, USA), respectively. Ultrapure water was produced using a Millipore water purification system. Ethanol was used for dissolve the antioxidant agent in the lipid peroxidation protocol, whilst acetone and n-hexane were used in chlorophylls analysis.

A 304 SS mesh with a pore size
of 0.5 mm² and a filament diameter of 270 μm was
used as a substrate. It measured 2 × 3 cm² in size.
The substrates were cleaned with deionized water, sanded with SiC
P180 abrasive paper, and rinsed again with water. Then, the substrates
were etched for 10 min using a 2 M FeCl₃ solution containing
deionized water, 37% HCl (w/w), and 5% H₂O₂ (w/w)
at a ratio of 15:1:1 (all reactants purchased from Scharlau). The
meshes were then rinsed in water and dried in an oven at 55 °C
for 1 h. After that, the etched substrates were immersed for 20 min
in a solution containing 0.15 M extra pure lauric acid (99.8% from
Scharlau) in 96% v/v absolute ethanol (synthesis grade from Scharlau)
at 60 °C. Finally, the treated substrates were cleaned with absolute
ethanol and dried in an oven at 55 °C for 45 min.

All reactants used were commercially available: hexadecylamine (HDA) 98% purity, oleic acid (OA) 99% purity, iron pentacarbonyl, dibenzyl ether 98% purity, ethyl acetate (99.5% purity), tetramethylammonium hydroxide (TMAH) solution in water (25% wt), and benzyl ether (98% purity) were purchased from Sigma-Aldrich Saint Louis, MO, USA and were used as received. Absolute ethanol and n-hexane 99% reagent grade were supplied by Scharlau, Cham Germany.

Genomic DNA extraction was carried out following the optimized protocol by our group [19] . 10⁸–10⁹ cells from control, UV irradiated, H₂O₂ treated and cryopreserved samples were resuspended to a total volume of 700 µl of extraction buffer (10 mM Tris-HCl, pH 8.0; 100 mM ethylenediaminetetraacetic acid (EDTA), pH 8.0; 0.5% (v/v) sodium dodecyl sulfate (SDS)), supplemented with 0.5 µl proteinase K (1 mg/ml). The samples were incubated overnight at 56°C in a shaking temperature-controlled bath (P-selecta, Spain). To eliminate cell components, one volume (700 µl) of phenol-chloroform-isoamyl alcohol mixture (25∶24∶1) was added to each sample. After shaking vigorously for 5 min and centrifuging at 12000×g for 5 min at 4°C, the aqueous phase was immediately collected. This step was repeated twice. Then, the aqueous phase was washed twice with chloroform and DNA precipitation was carried out using absolute ethanol (Scharlau, Spain). Finally, the DNA pellet was resuspended in 30 µl of TE buffer (1 M Tris-HCl, pH 8.0; 0.5 M EDTA) following the indications of Cartón et al., [19] .
DNA quantity and purity were determined spectrophotometrically at 260 nm (Nanodrop 1000, Thermo Scientific). Only high purity DNA (A₂₆₀/A₂₈₀ >1.8) was used for the qPCR assays.

Curcumin powder, low-molecular weight chitosan (75–85% deacetylated), Tween 80, sodium tripolyphosphate (TPP), sucrose, and sodium hydroxide (NaOH) were purchased from Sigma-Aldrich (St Louis, MO, USA). Acetic acid, ethyl acetate, and methanol were purchased from AMRESCO (Radnor, PA, USA). Absolute ethanol was purchased from Scharlau (Barcelona, Spain). All reagents utilised in this study were of analytical grade, except methanol, which was high-performance liquid chromatography (HPLC) grade. CrFK cells and FIPV strain WSU 79-1146 were purchased from ATCC (Manassas, VA, USA). Minimum essential medium (MEM) containing Earle's salts and L-glutamine, foetal bovine serum, nonessential amino acid (NEAA), penicillin (100 U/mL), streptomycin (100 μg/mL), phosphate-buffered saline (PBS), TryPLE™, trypan blue, dimethyl sulfoxide (DMSO), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Gold (III) acetate (Alfa Aesar, 99.9%), Palladium (II) acetate (Acros Organics, 47.5% Pd), Platinum (II) acetylacetonate (Acros Organics, 98%) were used as metal precursors. Titanium (IV) fluoride (TiF₄, 99%), titanium (IV) chloride (TiCl₄, 99%), 1-octadecene (90%) (1-ODE), 1-octadecanol (1-ODOL, 97%). Oleylamine (OLAM, 70%) and oleic acid (OLAC, 90%) were obtained from Sigma-Aldrich. Absolute ethanol was obtained from Scharlau. Milli-Q water was routinely used.

Triblock copolymer Pluronic P123 ((PEO)₂₀(PPO)₇₀(PEO)₂₀), tetraethyl orthosilicate (TEOS, Si(OC₂H₅)₄, 98%), ethylenediamine (EDA, C₂H₄ (NH₂)₂, ≥ 99%), hydrofluoric acid (HF, 40% in water), and magnesium nitrate hexahydrate (Mg(NO₃)₂.6 H₂O, 99%) were purchased from Sigma-Aldrich. Hexadecyltrimethyl-ammonium bromide (CTAB, CH₃(CH₂)₁₅N(Br)(CH₃)₃, 99%), and hydrochloric acid (HCl, 36.5%) were supplied from MERCK. Absolute ethanol (C₂H₅OH, 99%), sodium hydroxide (NaOH ≥ 99.0%),and 1000 PPM Magnesium standard solution for Atomic Absorption spectrometer were obtained from Scharlau. Carbon tetrachloride (CTC, CCl₄, ≥ 99.5%) was purchased from Pubchem. The materials were used as received without additional purification. The certified 10/90 vol% CO₂ /N₂ pre-mixed gas cylinder was purchased from ETICO gas.