Cell Line Maintenance and Validation Protocol

Small-molecule inhibitors were obtained from SelleckChem. siRNAs were obtained from GE Dharmacon. Cell lines were obtained from American Type Culture Collection (ATCC) and European Collection of Cell Cultures (ECACC) in 2010-2011 and maintained according to the supplier’s instructions as described in (19 (link),47 (link)). The MCF10A CDH1^-/- and MCF10A CDH1^+/+ isogenic cell lines, were obtained in 2014 from Sigma Aldrich, and maintained according to the supplier’s instructions. At sixth-monthly intervals and prior to storage, the identity of each cell line was confirmed by short tandem repeat (STR) profiling of 10 loci using the GenePrint 10 system (Promega). At monthly intervals, mycoplasma testing of cell cultures was carried out using MycoAlert Mycoplasma Detection Kit (Lonza).

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Bajrami I., Marlow R., van de Ven M., Brough R., Pemberton H.N., Frankum J., Song F.F., Rafiq R., Konde A., Krastev D.B., Menon M., Campbell J., Gulati A., Kumar R., Pettitt S.J., Gurden M.D., Cardenosa M.L., Chong I., Gazinska P., Wallberg F., Sawyer E.J., Martin L.A., Dowsett M., Linardopoulos S., Natrajan R., Ryan C.J., Derksen P.W., Jonkers J., Tutt A.N., Ashworth A, & Lord C.J. (2018). E-cadherin/ROS1 inhibitor synthetic lethality in breast cancer. Cancer discovery, 8(4), 498-515.

Publication 2018

Small-molecule inhibitors siRNAs MCF10A CDH1 GenePrint 10 system MycoAlert Mycoplasma Detection Kit

Cdh1 Cell cultures Cell line European Inhibitors Mycoplasma Promega Short tandem repeat Sirnas

Corresponding Organization : UCSF Helen Diller Family Comprehensive Cancer Center

Other organizations : King's College London, The Netherlands Cancer Institute, INCLIVA Health Research Institute, Universitat de València, Guy's Hospital, University College Dublin, University Medical Center Utrecht, Cancer Genomics Centre

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Variable analysis

independent variables

Small-molecule inhibitors obtained from SelleckChem
SiRNAs obtained from GE Dharmacon

dependent variables

Measured outcomes

control variables

Cell lines obtained from American Type Culture Collection (ATCC) and European Collection of Cell Cultures (ECACC) in 2010-2011 and maintained according to the supplier's instructions
MCF10A CDH1+/+ isogenic cell line obtained in 2014 from Sigma Aldrich and maintained according to the supplier's instructions
Mycoplasma testing of cell cultures carried out at monthly intervals using MycoAlert Mycoplasma Detection Kit (Lonza)

controls

Positive control: MCF10A CDH1+/+ isogenic cell line
Negative control: Not explicitly mentioned

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