09 mM CaCl2, 0 081 mM MgSO4∙7H2O, 3 μM H3BO3, 2 1 μM MnCl2∙4H2O,

09 mM CaCl2, 0.081 mM MgSO4∙7H2O, 3 μM H3BO3, 2.1 μM MnCl2∙4H2O, 1 μM Na2EDTA∙2H2O, 0.6 μM FeCl3∙6H2O, 0.03 μM

NaMoO4∙2H2O , 0.025 μM ZnCl2, , 0.01 NCT-501 cell line μM CoCl2∙6H2O, 0.07 nM CuCl2∙2H2O in double deionized water. Cyanidioschyzon merolae 10D was acquired from the Microbial Culture Collection of the National Institute for Environmental Studies (Tsukuba, Japan). Cyanidioschyzon was propagated using a Cyanidium medium [37] composed of 9.85 mM (NH4)2SO4, 2.06 mM K2HPO4, 1.01 mM MgSO4∙7H2O, 0.67 mM CaCl2, 13 μM Na2EDTA, 3.0 μM H3BO3, 2.2 μM FeCl3 .6H2O, 1.2 μM MnCl2∙4H2O, 0.32 μM CuSO4∙5H2O, 0.22 μM ZnSO4∙7H2O, 0.12 μM Na2MoO4 and 0.05 μM CoCl2 .6H2O in double deionized water. The medium was adjusted to pH 3.5 with HCl. Synechococcus leopoliensis (UTEX 2434), a cyanobacteria species, was obtained from the Culture Collection of Algae, University of Texas at Austin. Cells were grown in medium using 50X Cyanobacteria BG-11 Freshwater Solution (Sigma Aldrich, catalogue # C3061) [68] that was diluted to 1X in double deionized water to final concentrations of: 17.65 mM NaNO3, 0.3 mM MgSO4∙7H2O, 0.24 mM CaCl2∙2H2O, 0.18 mM K2HPO4, 46.0 μM H3BO3, 31 μM citric acid, 21 μM ferric ammonium citrate, 9.1 μM MnCl2∙4H2O, 2.8 μM MnNa2EDTA, 1.7 μM NaMoO4∙2H2O, 0.77 μM ZnSO4∙7H2O, 0.32 μM CuSO4∙5H2O, 0.17 μM Co(NO3)2∙6H2O.

All chemicals were obtained from https://www.selleckchem.com/products/gm6001.html Sigma-Aldrich (Oakville, Canada) or Fisher Scientific (Ottawa, Canada). Ferrostatin-1 chemical structure Synechococcus and Chlamydomonas were grown in 1.0 L of their respective media in 1.5 L Pyrex Lck glass cylindrical bioreactors under fluorescent lighting of 150 μE /m2/s at 28°C. Cells were kept suspended by aerating at a 1 L per min flow rate. Cyanidioschyzon was grown similarly except that the temperature was maintained at 45°C [53]. Cell treatments The effect of sulfur nutrition on heavy metal resistance and biotransformation was investigated by exposing each species to supplemental sulfur treatments. Supplemental sulfur was provided in the form of sulfate, sulfite or cysteine. Sulfate and sulfite were added as K2SO4 and K2SO3,

respectively, at ten-fold the amount of sulfur equivalents in the original media and the L-cysteine treatments were supplemented to twice the original amount of sulfur equivalents in the media. Experimental treatments included 1) no additional sulfur containing compounds, 2) additional sulfur containing compound, and 3) additional sulfur containing compound both before (pre-fed) and during the treatment period (plus). All treatments were performed in 100 mL of medium in 150 mL glass plant tissue culture vessels with translucent magenta B-caps obtained from Sigma-Aldrich (Oakville, Canada). Continuous fluorescent illumination was at 150 μE / m2/ s with 120 rpm rotary shaking. Culturing temperatures were 27°C for Synechococcus and Chlamydomonas, and 45°C for Cyanidioschyzon. The initial cell density for all cultures was O.D.665 = 0.1. These were grown to an O.D.665 = 1.

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