β estradiol

Demethoxycurcumin (DMC) was provided by Guangzhou Fans Biotechnology Co., Ltd. (Guangzhou, China). Alamethicin, D-saccharic-1,4-lactone, magnesium chloride (MgCl₂), MK571, Ko143, propofol, and uridine diphosphate glucuronic acid (UDPGA) were purchased from Sigma-Aldrich (St. Louis, MO). Human UGT1A1 and individual human liver microsomes (iHLM, n = 12) were obtained from Corning Biosciences (New York, USA). UGT1A1-overexpressing HeLa cells (HeLa1A1 cells) were established as described in a previous study [22 (link)] and provided by Prof. Baojian Wu, who works in Jinan University in Guangzhou of China. β-Estradiol and β-Estradiol-3-O-glucuronide were purchased from Toronto Research Chemicals (North York, ON, Canada). The anti-UGT1A1 antibody was purchased from BD Biosciences (Woburn, MA). The anti-BCRP (catalog number TA322704), anti-MRP1 (catalog number TA309559), anti-MRP2 (catalog number TA313641), anti-MRP3 (catalog number TA314800), and anti-MRP4 (catalog number TA327332) antibodies were purchased from OriGene Technologies (Rockville, MD). All other chemicals and reagents (analytical grade or better) were commercially available.

Yeast strains used in this study are described in Supplementary Table 2. Yeast strains were cultured at 30 °C in yeast extract peptone dextrose (YPD) (10 g/l yeast extract, 20 g/l peptone, 2% (w/v) glucose) or synthetic complete (SC) medium with two or three amino acids dropped out to select for the maintenance of plasmids carrying corresponding auxotrophic markers. The synthetic yeast strains used for genome rearrangement via GCE-SCRaMbLE were grown in SC–His–Leu medium supplemented with various concentrations of OMeY (Sigma-Aldrich, Cat number: 158259). The synthetic yeast strains used for fluorescence assay were grown in SC–His–Leu–Ura medium supplemented with OMeY or β-estradiol (Toronto research chemicals, Cat number: E888000). The isolated SCRaMbLEd strains for genome extraction and sequencing were grown in YPD to stationary phase. The synthetic diploid strains were constructed by mating the yeast strain bearing multiple synthetic chromosomes (synII, synIII, synVI, and synIXR) to BY4741 and the ring_synII synthetic strain to BY4742. The diploid strains were selected and confirmed by polymerase chain reaction (PCR) using specific MATa/α primers listed in Supplementary Table 3.