Animal procedures were performed according to a protocol approved by Northeastern University, Institutional Animal Care and Use Committee (NUIACUC). Five to six weeks old female nu/nu (athymic) mice, strain CrTac:NCr-Foxn1nu, weighing approximately 20 g, were purchased from Taconic Biosciences, Inc. (Hudson, NY). The animals were allowed to acclimate for at least 72 h prior to any experimentation, raised under specific pathogen-free conditions, kept in individually ventilated cage racks and supplied with sterile rodent pellets and water ad libitum. Mice were housed under a 12 h light/dark cycle.
For A549-WT and A549-DDP tumor model development, mice were injected subcutaneously with 3×106 cells in a mixture of 50 µL DMEM/F12 medium and 50 µL Matrigel under mild anesthesia on their right flank. Tumor volume was calculated by the modified ellipsoid formula: tumor volume = 1/2(length × width2). When tumors grew to approximately 200 ± 20 mm3 in volume the animals were randomized into groups to yield even distribution of tumor sizes. Sample size was determined by power analysis using the software G*Power. Each animal was identified with an ear tag which allowed us to perform blind experiments in order to reduce bias in animal selection and outcome assessment. The animals were monitored daily for food/water intake, body weight and any physical signs of discomfort.
For A549-WT and A549-DDP tumor model development, mice were injected subcutaneously with 3×106 cells in a mixture of 50 µL DMEM/F12 medium and 50 µL Matrigel under mild anesthesia on their right flank. Tumor volume was calculated by the modified ellipsoid formula: tumor volume = 1/2(length × width2). When tumors grew to approximately 200 ± 20 mm3 in volume the animals were randomized into groups to yield even distribution of tumor sizes. Sample size was determined by power analysis using the software G*Power. Each animal was identified with an ear tag which allowed us to perform blind experiments in order to reduce bias in animal selection and outcome assessment. The animals were monitored daily for food/water intake, body weight and any physical signs of discomfort.