Wild-type H1975 control (siCtrl) or CAR knockdown (siCAR-1 and siCAR-2) cell lines (3 × 106) were injected subcutaneously into the two posterior flanks of BALB/c nude mice (Charles River Laboratories). Nine 5-week-old female mice in total were used for control cell lines and five for each siRNA. Mice were followed daily, tumors were measured with a caliper in long and short axes, and volume was determined on the basis of the equation 0.4 × A × B2 (where A is the long axis of the tumor and B is the short axis of the tumor). At day 14 after injection, mice were culled using CO2 and tumors were removed aseptically with dissecting scissors and weighed. All animals were maintained under specific pathogen–free conditions and handled in accordance with the Institutional Committees on Animal Welfare of the UK Home Office Animals (Scientific Procedures) Act 1986. All animal experiments were approved by the Ethical Review Process Committee at King’s College London and carried out under license from the Home Office, UK.

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