Balb c nu mice

Four-week old male BALB/c-nu mice were obtained from Charles River Laboratories, Japan, and acclimated for one week. Mice were raised in 22 ± 3 °C with saturated humidity and a standard light/dark cycle. Plenty of food and water was guaranteed.
The same amounts of control and 2FF pre-treated HepG2 cells (4 × 10⁶) were injected subcutaneously into the left and right flanks of each mouse, respectively. Based on the inhibitory effect of 2FF on fucosylation lasted for 7 days at least (Fig. 1C), 100 μl of the DMEM solution containing with 2FF at 100 μM were directly injected into each tumor tissue from three directions after the inoculation for 7 days, and once a week for 3 weeks. All the mice were euthanized at 25 days after the first injection of 2FF. Tumor tissues were isolated, photographed and then weighed. And the same amounts of tumor tissues were homogenized for lectin blot with AAL and Western blot with several indicated antibodies. Measurement of tumor sizes began on the fifth day, and sizes were monitored every 5 days by measuring the tumor length and width with a vernier caliper. Tumor volumes were estimated according to the following formula: volume (mm³) = (L × W²)/2, where L and W are the length and width of the tumor tissues. All experiments were performed according to protocols approved by the Tohoku Medical and Pharmaceutical University Research Ethics Board.

Sixteen 4-week-old female BALB/c-nu mice were purchased from Charles River Laboratories and were raised in the specific pathogen free animal room at Chengdu Medical College Research Center. The mice were injected subcutaneously with 5 × 10⁶ 22RV1 cells to establish a xenograft tumor model. After 3 days of acclimatization feeding, all the experimental mice were randomly divided into the experimental group and the control group, with eight mice in each group. During the experiment, each group was administered the control diet, while the experimental group was additionally fed with genistein (100 mg/kg.bw/day) via gavage feeding.
The volume of tumors in the mice was measured every 3 days using vernier callipers, with measurements taken in two vertical directions. The following formula was used to estimate the tumor volume: volume = 0.52 × (length) × 2 (width). After 30 days of breeding, the mice were executed, and xenograft tumors were collected for weighing, immunohistochemistry, and ki-67 staining.

Laboratory animal handling and experimental procedures were performed in accordance with the requirements of Provisions and General Recommendation of Chinese Experimental Animals Administration Legislation. The research project was examined and certified by the Ethics Committee of the General Hospital of Ningxia Medical University. PC-9-GR cells (1 × 10⁶ cells · 0.1 ml per mouse) were inoculated subcutaneously into the right front axilla of female athymic BALB-c/nu mice at 5 to 6 weeks of age (Charles River, Beijing, China). Treatment of 6 mice per group was started when the tumors had reached a volume of 150 to 200 mm³ with vehicle control, gefitinib (50 mg/kg, 5 days a week), AT-101 (35 mg/kg, 5 days a week), or gefitinib plus AT-101. Both drugs were orally administered. Tumor volume was determined from caliper measurements of tumor length (L) and width (W) according to the formula LW²/2. Both tumor size and body weight were measured twice per week. Immunohistochemical analysis was performed on formalin-fixed, paraffin-embedded tissue sections staining for Ki-67 (Cell Signaling Technology, Beverly, MA, USA) and cleaved caspase-3 (Santa Cruz, CA, USA).

Four to five-week-old female nude (BALB/c-nu) mice were purchased from Charles River (Wilmington, MA, USA). All animal experiments were conducted according to the protocol approved by the Animal Care and Use Committee of Kyoto University (IACUC): Title of the protocol, “Chemosensitivity studies of gastrointestinal cancers using patient-derived tumor xenografts.” Approval No. 14546, 15091, 16047, 16654, 17086, 18080, and 19601 (2014–2019).

For assessment of UMI-77 anti-tumour activity in vivo, 3 million MDA-MB-468 breast cancer cells were injected bilaterally into the inguinal mammary fat pads in 1:1 PBS:matrigel mix into 8-week BALB/c-Nu female mice (Charles River, UK). Treatment commenced 2 weeks after injection and UMI-77 was administered by intraperitoneal injection at 60 mg/kg in a regime of 5 daily doses followed by 2 rest days. For in vivo use UMI-77 was dissolved in 5% DMSO/30% PEG300/ 65% dd H₂0. Tumour growth was monitored by caliper measurement three times per week and volume calculated using the equation ([length × width²]2). Graphs represent average of three weekly measurements relative to tumour volume at commencement of treatment. Tumours were harvested after 4 weeks of treatment.
MCL-1 knockdown was achieved using a pool of prevalidated siRNA to human MCL-1 s8583 (Ambion/Life Technologies, UK) at 5 nM concentration or non-targeting control siRNA and nucleofection using Amaxa kit (Lonza, UK) according to the manufacturer's protocol. For orthologous transplantation assay of siRNA-treated MDA-MB-468 cells, 3 million siMcl1 or siSCR treated cells in a 1:1 PBS:matrigel mix were injected bilaterally into the inguinal mammary fat pads of 6 week female BALB/c-Nu mice (Charles River, UK) 18 h after nucleofection.