Lncap and pc3 cells

Human prostate carcinoma LNCaP and PC3 cells were obtained from American Type Culture Collection (Manassas, VA). Cells were cultured in RPMI 1640 with 10% fetal bovine serum (Hyclone, Logan, UT) under standard culture conditions (37 °C, 95% humidified air and 5% CO₂). Cells were plated at a density of 5 × 10³ cells/cm² plates under standard culture conditions. After 24 h, cells were treated with either DMSO alone (control), or different doses of Nintedanib dissolved in DMSO (concentration did not exceed 0.1% in any treatment (v/v)). At the end of treatment time (72 h), cells were collected after brief trypsinization, washed with PBS, and then stained with Trypan blue. The total cell number (colorless and blue stained) and dead cells (blue stained) were counted under light microscope using hemocytometer.

CP2 and SP1 cells were derived from genetically modified mouse prostate cancer models that represent activation of β-catenin and inactivation of Sprouty2 along with the loss of Pten tumour-suppressor protein, respectively.^{10 (link),11 (link)} Details of the CP2 (RRID:CVCL_VQ85) and SP1 (RRID:CVCL_VQ86) cell lines have been deposited on the RRID Portal (https://scicrunch.org/resources/). Cells were grown in DMEM supplemented with 10% foetal bovine serum (FBS) and 2 mM l-glutamine. LNCaP and PC3 cells were obtained from American Type Culture Collection and were grown in RPMI supplemented with 10% FBS and 2 mM l-glutamine. RPE1 cell lines stably expressing H2B-RFP, GFP-tubulin or EB3-GFP were maintained in the DMEM/F-12 medium supplemented with 10% FCS, 2.3 g/l sodium bicarbonate, 100 U/ml penicillin, 100 μg/ml streptomycin and 500 µg/ml geneticin. Cell lines were authenticated by LCG standards or in-house using Promega GenePrint 10 Kit. All cell lines used were routinely tested every 6 months for mycoplasma using an in-house MycoAlert™ Mycoplasma Detection Kit (Lonza, Switzerland), according to the manufacturer’s instructions. RPE1 cells were tested monthly for mycoplasma using a MycoSensor PCR Assay Kit (Agilent Technologies, USA).

LNCaP and PC3 cells were purchased from the American Type Culture Collection (Manassas, Virginia, USA) and cultured under typical conditions in RPMI 1640 (Invitrogen) supplemented with 10% fetal bovine serum (FBS). For androgen‐depleted conditions, cells were cultured in phenol red‐free RPMI 1640 (Invitrogen) supplemented with 10% charcoal‐stripped fetal bovine serum (CSFBS) (Hyclone). Cell numbers were counted using a Countess II (Invitrogen).