Cyp1A- och ABC-transportöruttryck och funktion i PLHC-1cellinje efter exponering för mikrocystin-LR och benso (a)pyren

The data were collected through laboratory experiments conducted at the Department of Biological and Environmental Sciences, University of Gothenburg. All experiments took place in 2022, except for the half-maximal inhibition concentration (IC50) assays for benzo[a]pyrene (BaP) on CYP1A-mediated EROD activity in PLHC-1 cells pre-exposed to BNF (performed in July 2024). The PLHC-1 cell line (ATCC® CRL-2406TM) was obtained from LGC Standards (Borås, Sweden). Cells were cultured in tightly capped 75 cm² cell culture flasks with 25 mM HEPES-buffered MEM, pH 7.1, supplemented with 5% (v/v) FBS at 30 ºC. For each assay, cells were seeded as follows: - EROD and Rh123 accumulation assay: 48-well plates, 6 × 10⁵ cells/mL in 500 µL per well. - Cytotoxicity Analyses: 96-well plates, 5 × 10⁵ cells/mL in 200 µL per well. - mRNA Expression Analyses: 6-well plates, 2 × 10⁶ cells/mL in 2 mL per well. After a 24-hour incubation post-seeding, media were removed from each well, and cells were rinsed with 10 mM Na-phosphate buffer, pH 7.4, containing 0.9% (w/v) NaCl (PBS). Cells were then exposed to DMSO (control), MCLR, BaP, or their mixture. EROD activity, Rh123 accumulation, cytotoxicity, and mRNA expression were measured on separate plates, with each type of measurement conducted on different days. Data were recorded using a VICTORTM 1420 Multilabel Counter (Wallac Sverige AB) for EROD, cytotoxicity, and Rh123 assays, and an iCYCLER instrument (Bio-Rad) for qPCR.