Cuso4

Susceptibility toward copper (Cu), zinc (Zn), cadmium (Cd), and mercury (Hg) was tested, as previously described (Sharifi et al., 2015 ; Capps et al., 2020 (link)). Briefly, overnight cultures were spotted onto Mueller–Hinton agar (Oxoid) supplemented with different concentrations (0.05 to 40 mM) of ZnCl₂ (Carlo Erba), HgCl₂ (Sigma Aldrich), and CdSO₄ (Sigma Aldrich) resuspended in distilled water, and CuSO4 (Merck Millipore) adjusted to pH 7.2 with 1 M NaOH. After 24 to 48 h of incubation at 37°C, the plates were visually inspected for bacterial growth on the spots.

E. coli was grown overnight at 37 °C, either in 2xYT liquid media (16.0 g tryptone, 10.0 g yeast extract, 5.0 g NaCl per 1 l of media) or on LB solid media (10.0 g tryptone, 5.0 g yeast extract, 5.0 g NaCl, 15.0 g agar per 1 l of media), supplemented with 100 mg/l ampicillin.
S. cerevisiae was grown at 30 °C/180 rpm in a chemically defined medium (6.70 g/l Difco Yeast nitrogen base without amino acids, 20.0 g/l glucose, 0.77 g/l MP Bio drop-out mixture, and 20.0 g/l agar for solid media). In the imaging experiments, Difco's YNB was replaced with Formedium's LoFlo YNB without amino acids, folic acid, and riboflavin (Formedium, Hunstanton, United Kingdom). In the experiments with copper, Difco's YNB was replaced with Formedium's YNB without amino acids and copper. Copper was supplemented as CuSO₄ (Merck, Germany). Strains expressing ymNeongreen [38 (link)], QUEEN-2 m [39 (link)], and sfpHlourin [40 (link)] were grown in -his medium, while strain expressing copper biosensor [36 (link)] was grown in -ura medium.

PAPC (850459C) and DPPC (850355) were purchased from Avanti Polar lipids and stored in chloroform at −80°C. POVPC (10031), KOdiAPC (62945), and PGPC (10044) were all purchased from Cayman Chemicals. Lipids were stored at 10 mg/ml in EtOH at −20°C. The lipids EC, PECPC, EI, PEIPC, 15d-PGJ2, and 15d-PGJ2PC were synthesized as described (Egger et al, 2013 (link), 2014 (link)). All lipids were stored at −80°C under nitrogen atmosphere. For experimental purposes, lipids were dissolved in DMSO to 20–50 mM in glass flasks (Carl Roth GmbH + Co) and then further diluted in pure RPMI-1640 medium before addition to cell suspensions. PAPC were oxidized by air exposure, with 5–10 μM iron(II)sulfate (Sigma, F8633-250G), or with 5–10 μM CuSO₄ (Merck, 1.02790.0250) as oxidizing agents for various time intervals. Metal-catalyzed oxidation was performed in sterile PBS at 37°C in glass flasks using a rotary wheel set at 20 rpm/min.

The SS mesh (SS316, 325 × 2,300, McMASTER-CARR) was cut into 0.2–1 cm_geo² pieces and then spot welded with Cu wire (≥99.98%, Goodfellow) for electrical connection and used as WE. Two Pt mesh (1.5 cm_geo², 99.9%, Goodfellow) electrodes were connected and used as a split CE. The WE was located in the middle of the two Pt meshes. Prior to electrodeposition, the WE was dipped in 0.06 M HCl (VWR Chemicals) and rinsed with Milli-Q water and ethanol. The electrolyte was made of 0.4 M CuSO₄ (98%, Merck) in 1.5 M H₂SO₄ (99.999%, Sigma Aldrich). A constant current of −2 A with applied time ranging from 15 s to 7 min was set for the porous Cu deposition on SS mesh. The electrode was cleaned with Milli-Q water and ethanol several times and dried under vacuum after electrodeposition. The excess deposited Cu on the Cu wire and the edge of SS mesh were removed to keep the constant geometric area of the WE at different experimental conditions (Figure S29). All the WEs were stored in an Ar glovebox before use.

Roots of Bistorta amplexicaulis plant was collected from Ghangachoti Bagh Azad Kashmir. Chemicals including AgNO₃, NaNO₃, KCl, CaCl₂, SnCl₂, BaCl₂·2H₂O, HgCl₂, PbCl₂, CuSO₄, Mg(NO₃)₂, Ni(NO₃)₂·6H₂O, AlCl₃, and CuSO₄·5H₂O were purchased from chemical companies i.e., Merck or Sigma Aldrich and used without any further pretreatment. A calculated amount of AgNO₃ and each metal salt were added in distilled water to prepare a stock solution.