Balb c nude mice

Six-week-old female BALB/c nude mice and 8-week-old female ICR mice were purchased from Nara Biotech (Seoul, Korea). All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of Yonsei University, Mirae Campus (YWCI-201706-011-03) and performed in accordance with the school guidelines and regulations.

Animal use complied with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and was approved by the Institutional Animal Care and Use Committee at Sejong University. Five-week-old immune-deficient BalB/c nude mice (Nara Biotech, Seoul, Korea) were group housed in pressurized ventilated cages under a 12:12 light-dark lighting schedule with free access to food and water in an SPF facility. Transfected HCT116 cells (1 × 10⁷) were injected into mice subcutaneously. Tumor growth was monitored periodically and measured using calipers. Tumor volumes were calculated by the formula: Tumor volume = width² × length × 1/2. The weight of the tumor was measured after sacrificing the animal.

Five-week-old female BALB/c nude mice were purchased from Nara Biotech (Seoul, Korea) and housed under specific pathogen-free facility under constant conditions (12 h light-dark cycle at 22 ± 1°C and 55 ± 5% humidity). All animal experiments were approved by the Animal Care and Use Committee of the Korea Institute of Oriental Medicine (KIOM, Daejeon, Korea) with reference number #14–040 and performed according to the guidelines of the Animal Care and Use Committee at KIOM.

All procedures were approved by the Institutional Animal Care and Use Committee of the Kangwon National University (#KIACUC-11-0006) and carried out in compliance with the ARRIVE guidelines (https://arriveguidelines.org/). Female BALB/c nude mice at 6 weeks of age were obtained from Nara Biotech (South Korea). Five to seven mice were allocated to each group and then injected subcutaneously in each flank with 3 × 10⁶ MEFs in 100 μl of PBS. Tumor volumes were measured with an electronic caliper every 2–3 days and calculated with the formula x² x y/2, where x is the width and y is the length^{43 (link)}. All experiments were performed in accordance with relevant guidelines and regulations.

In total, 5 × 10⁵ Molm-13 cells were prepared for each mouse and resuspended in 50 μL of PBS. Cells were mixed with 50 μL of Matrigel matrix (354234, BD Sciences). The mixtures of cells and matrigels were subcutaneously injected in the flank of 6-week-old female Balb/c nude mice (Narabiotech, Seoul, Korea). After one week, mice with xenograft tumours were inoculated with vehicle or birinapant (2 mg/kg) plus emricasan (1 mg/kg) by i.p. injection in every two days. Tumour growth was measured as described previously [35 (link)]. After 18 days from tumour inoculation, mice were sacrificed, and tumour volumes and masses were determined as described previously [35 (link)]. We analysed six mice for each group. Animal experiments were approved by the Institutional Animal Care and Use Committee of the Laboratory Animal Research at Yonsei University (IACUC-A-201905-916-01). Xenograft analyses were not randomized. The investigators were not blinded to allocation during experiments and outcome assessments.