Sodium dodecyl sulfate (sds)

Fly heads were homogenized using an ultrasonicator at 4 °C for 6 min at 40 kHz in 3 µL of total protein extraction kit TM buffer containing 0.1% SDS (Millipore). The homogenized lysates were centrifuged at 17,000 × g for 1 min at RT to remove the debris. The supernatant was collected and centrifuged again under the same conditions to collect the fresh supernatant. Four-week-old mnd2/mnd2 mice, mnd2/+ mice and aged match controls were sacrificed, and the brain tissues were chopped into small pieces, followed by the addition of 1 mL of total protein extraction kit TM buffer containing 0.1% SDS (Millipore) to 0.2 g of brain samples. Samples were homogenized, and total proteins were isolated as described above.

CD measurements for secondary structure estimation were performed on a Jasco J 715 Spectropolarimeter (Jasco, Gross-Umstadt, Germany) at room temperature using quartz cuvettes with an optical path length of 0.02 cm. The CD spectra were measured between 260 nm and 180 nm with a 0.2 nm step resolution. To improve accuracy, 3 scans were accumulated and averaged. The peptides were dissolved in 20 mM phosphate-buffered saline (PBS) (20 mM NaPi, without NaCl, pH 7.4). Spectra were measured in the absence and presence of 20 mM POPS, 20 mM POPC and 0.5 µM Dodecylphosphocholine (DPC) (Avanti Polar Lipids, Alabaster, USA) and 0.5 µM sodium dodecyl sulfate (SDS) (Sigma Life Science, Darmstadt, Germany), with POPS and SDS mimicking cancer, POPC and DPC healthy mammalian membranes, respectively. Liposomes were prepared as described above. The respective peptide to surfactant molar ratios were 25:1. The background signals were subtracted after the measurements. Percentage secondary structure calculations were conducted using Spectra Secondary structure estimation [78 (link)].

Gold nanoparticles (AuNPs) of 13 nm diameter was prepared as reported in previous work [24 (link)]. Before conjugation, detecting probe (DP; Bio Basic Inc., Ontario, Canada; sequence: 5′-SH-C6-ATC ATC GA AGT GGC TTC A-3′) was prepared by adding 500 mM acetate buffer (pH 4.68, which consist of 1 M sodium acetate (Ajax Finechem Pty. Ltd., NSW, Australia) and 0.21 M acetic acid (Fisher Scientific) to the lyophilized DP and topped up to 100 μM with distilled water. AuNP–DP conjugates were prepared by adding 6 μL of 100 μM detecting probe (DP) to 1 mL AuNPs solution and left for 16 h for conjugation. 1% sodium dodecyl sulfate (SDS) (Sigma Aldrich) and 2 M NaCl (Merck) was then added to the solution to achieve a final concentration of 0.0% SDS and 0.16 M NaCl in the solution. The solution was then allowed to age for at least 24 h before centrifugation at 14,000 g for 25 min. Following centrifugation, the supernatant was removed while the pellet was resuspended in 67 μL eluent buffer, which comprised of 5% bovine serum albumin (w/v; AMRESCO, Solon, OH, USA), 0.25% Tween-20 (w/v; Sigma Aldrich), 10% sucrose (w/v; Ajax Finechem Pty. Ltd.) and 20 nM trisodium phosphate (Sigma Aldrich). The resulting AuNP–DP conjugate solution was kept at 4 °C for future use. We have characterized the AuNPs in our previous publication. Their average sizes are 13 nm [3 (link)].

Allium sativum (PUSA- AG 102) commonly known as “garlic” was obtained from IARI, New Delhi. Chemicals; trypsin (bovine pancreatic trypsin), Nα-benzoyl-DL-arginine-p-nitroanilide (BAPNA), phenylmethylsulfonyl fluoride (PMSF), Polyvinylpyrrolidone (PVP), acrylamide, bis-acrylamide, Tetramethylethylenediamine (TEMED), ammonium persulfate and Sodium Dodecyl Sulfate (SDS), acrylamide, bis-acrylamide, TEMED, ammonium persulfate and SDS were obtained from Sigma-Aldrich. All other reagents and chemicals used were of analytical grade.

All chemicals and reagents were of analytical grade and used without any further purification: zinc acetylacetonate hydrate, Zn(acac)₂ (Sigma-Aldrich, St. Louis, MO, USA, M = 263.61 g/mol), Oleylamine (C₁₈H₃₅NH₂), OAm (Merck, Rahway, NJ, USA, M = 267.493 g/mol), Sodium Dodecyl Sulfate (CH₃(CH₂)₁₁SO₄Na), SDS (Sigma-Aldrich, ≥99%, M = 288.38 g/mol), triethylene glycol, TrEG (Merck, ≥99%, M = 150.17 g/mol), diethyl ether (Merck, ≥99%, M = 74.12 g/mol), geraniol, trans-3,7-Dimethyl-2,6-octadien-1-ol (Sigma-Aldrich, 98%, M = 154.25 g/mol), Milli-Q water, phosphate buffer solution, PBS (Gibco by Life Technologies, Carlsbad, CA, USA, pH 7.2, 10X), potato dextrose agar, PDA (BD Difco, Franklin Lakes, NJ, USA, Luna^® Sensation SC Fungicide (Bayer Crop Science, Leverkusen, Germany, fluopyram 250 g/L and trifloxystrobin 250 g/L), and methanol (SD-Fine).