Balb cslc nu nu

Pathogen-free female nude mice (BALB/c Slc-nu/nu), aged 4 weeks and weighing 20–25 g, were purchased from Japan SLC, Inc. (Hamamatsu City, Shizuoka, Japan). Xenograft tumor models were established by implanting 100 μL of medium containing 1 × 10⁶ HCT116 cells subcutaneously in the right flank of each animal. The use of animals and the procedures employed in this study were approved by the Animal Research Committee of Nagoya City University (Project number: H29M-38).

All animal experimental procedures were approved by the Institutional Animal Care and Use Committee of JAIST and AIST. Female BALB/cSlc (4 weeks old; average weight, 16 g) and BALB/cSlc-nu/nu mice (4 weeks old; average weight, 18 g) were purchased from Japan SLC (Hamamatsu, Japan) and were housed in specific pathogen-free facilities with a 12-h light/12-h dark cycle and free access to food and water.
To generate tumour models, nude mice were subcutaneously inoculated in the flanks with equivalent number of cells (5 × 10⁵ cells for HT-29; 1 × 10⁶ cells for A549 and Colon-26) suspended in 100-μl aliquots of culture medium/Matrigel (Corning) mixture (v:v = 1:1). For in vivo tumour phototherapy experiments, mice were bilaterally implanted with either control or TRPV2 overexpressing derivative cells into flanks. For in vivo biodistribution analyses, both cell types were injected into opposite flanks of the same mouse. For in vivo drug and phototherapy combination experiments, only TRPV2-transfected A549 cells were injected into the right flank of mice. Tumour sizes were monitored using vernier calipers and tumour volumes were calculated as V = L × W²/2, where L and W denote lengths and widths of tumours, respectively. Tumour models were randomly divided and used in experiments when tumour volumes reached about 100 mm³.

The protocols for all animal studies were approved by Nagoya City University Center for Experimental Animal Science, and the mice were housed according to the guidelines of Nagoya City University for Animal Experiments. Female nude mice (BALB/c Slc-nu/nu), aged 7 weeks, were obtained from Japan SLC Inc. The mice were acclimatized for 2 weeks before the experiments, and were kept in individual cages with unrestricted access to food and water. All mice were maintained under specific pathogen-free conditions with a 12-h light/dark cycle. To prepare the xenograft models, HuCCT-1 cells were injected into the mouse flanks with 5 × 10⁶ cells in 100 μL of media. One day after implantation, the mice were randomly allocated into two groups. Two weeks after subcutaneous tumor transplantation, UA (20 mg/kg, 3 times a week) or dimethyl sulfoxide (DMSO; control) was administered by oral gavage, as in a previous study (20 (link)). The maximum tumor diameter (L) and the diameter at right angles to that axis (W) were measured using calipers twice a week, and the tumor volume was calculated according to the formula: (L×W2)/2. The weights of the mice were also recorded twice a week. The mice were sacrificed 35 days after the start of medication, and the transplanted tumors were excised and fixed in formalin or frozen in liquid nitrogen for protein lysate.

Immunodeficient mice (BALB/c Slc-nu/nu, Japan SLC, Inc.) were subcutaneously injected with 1 × 10⁶ cells suspended in 200 μl of serum-free McCoy’s 5A for one location. Tumors were monitored every 2 or 3 days, and then mice and tumor volumes were calculated using the following formula: 0.5 × L × W². The mice were sacrificed after 16 days with tumor volume less than 1 cm³ of monitoring. The mice used this study were housed in environmentally-controlled rooms of the animal experimentation facility at Osaka University and sacrificed under deep anesthesia by inhaling 4% isoflurane. Experiments were conducted under the applicable laws and guidelines for the care and use of laboratory animals in the Research Institute for Microbial Diseases, Osaka University, approved by the Animal Experiment Committee of the Research Institute for Microbial Disease, Osaka University.

Animal experimental protocols were approved by the Committee for Animal Research and Welfare of Gifu University. BALB/cSLC-nu/nu (nude) mice were obtained from Japan SLC (Hamamatsu, Japan). After a mouse had been killed, protein and RNA were extracted and used for Western blotting and real-time reverse transcription-PCR analysis, respectively. Details of the methods are given below in the Western blotting and Real-time reverse transcription-PCR sections.

All animal studies were approved by the Animal Care and Use Committee of Tohoku University. Male nude mice (BALB/cSlc-nu/nu, purchased from Japan SLC, Inc) were fed standard chow (CE-2, CLEA Japan Inc.) ad libitum in a temperature- and humidity-controlled environment with a 12 h light/12 h dark cycle (08:00–20:00) at constant temperature (23 °C). Transplantation of cultured cells was operated as previously reported^{70 (link)}. 3T3-L1 preadipocytes transduced with empty virus or SETD5 were cultured in differentiation medium for two days. Subsequently, cells were collected by scraping and resuspended in the basal medium containing 1 μg/ml insulin and mixed with 0.25% HydroMatrix Peptide Cell Culture Scaffold (Sigma-Aldrich). Cells (1.5–2 × 10⁶ cells) were injected into the subcutaneous region on the upper or lower back of nude mice at 6 weeks of age. Two weeks after injection, transplanted cells were isolated with skin and subjected to histological analysis.

Seven‐week‐old female BALB/c nude mice (BALB/cSlc‐nu/nu) were purchased from Japan SLC, Inc. All experimental protocols were approved by the Korea Research Institute of Bioscience and Biotechnology (KRIBB) Animal Care and Uses Committee. All of procedures were performed by accordance with the appropriate KRIBB biosafety guidelines and regulations. Mice were anaesthetized with 2% Avertin solution (0.1 ml/20 g body weight; Bayer, Leverkusen, Germany). The back of the mouse was sterilized using an alcohol swab and a sterile biopsy punch (6‐mm diameter) was used for making two wounds in their dorsal skin. Mice were randomized into the following four groups: a control group, which received 50 μl of 1 × 10⁶/each site unsorted cells at passage 5, EPCs group, which received 50 μl of 1 × 10⁶/each site EPCs, EPCs‐shCTL group, which received 50 μl of 1 × 10⁶/each site EPCs transduced the shCTL lentivirus, and EPCs‐shITGB1 group, which received 50 μl of 1 × 10⁶/each site EPCs transduced the shITGB1 lentivirus. The wound area was measured every day for 10 days after wounding. On the tenth day, mice were sacrificed, wound skin samples were collected, fixed in 4% paraformaldehyde, embedded, and sectioned for morphometric analysis and immunohistochemistry.