MCF-7 cells were treated with different compounds for 8 h to analyze differential gene expression

MCF-7 cells were grown in phenolred-free, high glucose DMEM medium, supplementent with 10% FBS and 1% Penicillin/Streptomycin. Cells were incubated at 37°C, 90% humidity, and 5% CO2. Cells were seeded 24 h before treatment. MCF-7 cells were treated for 8 h with the compounds of interest at a concentration of 25 µM. MCF-7 cells were further treated with 0.2% DMSO as control. Cells were harvested with phenolred-free trypsin and RNA extraction was performed with the InviTrap® Spin Cell RNA Mini Kit (Invitek Molecular GmbH, Berlin, Germany), according to the manufacturer's instruction. RNA sequencing was performed by StarSEQ GmbH, Mainz, Germany. RNA quality was verified with a 2100 Bioanalyzer system. The NEBNext© Ultra™ II Directional RNA Library Prep Kit (New England Biolabs, MA, USA) was used for mRNA preparation. Sequencing was carried out with the Illumina NextSeq 500™ system using 25 Mio paired-end reads (2 × 150 nt). Each sample was measured as biological duplicates.