De17 602e

PC-3M-luc cells and DU145 cells were cultured in EMEM (Lonza, Basel, Switzerland, 12–125) supplemented with 10% FCS, 1% L-glutamine (Lonza, BE17-605E) and 1% penicillin-streptomycin (Lonza, DE17-602E). RNAi knockdown was performed using Dharmafect 2 (Thermo Scientific, Waltham, MA, USA, T-2002-01) in PC-3M cells and Dharmafect 4 (Thermo Scientific, T-2004-01) in DU145 cells using 25 nM siRNA according to the manufacturer's instruction. For knockdown of LSD1 the following siRNAs were used: siCtrl: 5′-AACGTACGCGGAATACTTCGA-3′ and siLSD1 5′-acacaaggaaag cuagaagaa-3′. For others siCtrl 5′-GAAAGTCCTAGATCCACACGCAAAT-3′, siLPAR6 5′-UCAGCAUGGUGUUUGUGCUUGGGUU-3′, siPXN 5′-CATACCCAACTGGAAACCACACATA-3′, siTLN1 5′-CATTGTACTTGATACGGCCAGTGAT-3′, siZYX 5′-CAGGGAGAAGGTGAGCAGTATTGAT-3′ and siPIK3R2 5′-CCCTCAGGAAAGGCGGGAACAATA-3′. Dharmafect Duo (Thermo Scientific, T-2010-01) was used for plasmid transfection as described in the manual. Puromycin (Sigma, St Louis, MO, USA, P8733, 5 μg/ml) was administered to cells 24 h post transfection. 293T cells were cultured in DMEM supplemented with 10% FCS, 1% L-glutamine and 1% penicillin-streptomycin. Viral production was performed as described.^{12 (link)} PC-3M cells were infected with FU-GFP or FU-GFP-LSD1 and subjected to migration assay 24 h post transduction.

Adult, female (8 to 10 weeks old) C57BL/6J mice were purchased from Harlan (Horst, The Netherlands). The mice were housed in conventional pathogen-free conditions. All animal experiments were approved by the bioethics committee of the KU Leuven that follows international guidelines.
Methylcholanthrene-induced murine C57BL/6J syngeneic GL261 glioma cells were kindly provided by Dr. Eyüpoglu (University of Erlangen, Germany). The cells were maintained in DMEM supplemented with 10% heat-inactivated FCS (Sigma-Aldrich, F7524), 2 mM L-glutamine, 100 U/mL penicillin and 100 mg/mL streptomycin (both from Lonza, BE17-605E and DE17-602E, respectively). Cell morphology was evaluated by microscopic examination multiple times per week.

Murine melanoma B16.F10 (ATCC, CRL-6475) cancer cells were cultured in Dulbecco’s Modified Eagle medium (DMEM, Lonza), supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 IU/ml penicillin and 100 µg/ml streptomycin (DE17-602E, Lonza), and 4 mM L-Glutamine (BE17-605E, Lonza). Cancer cells were maintained as a monolayer at 37°C in a 5% CO₂ humidified atmosphere.