Mi se software

FITC-conjugated TGF-β2 was employed for fluorescence signal tracing, and was prepared as previously described. For in vivo experiments, 5 × 10⁵ P1- LbL-coated HFSCs loaded with FITC-conjugated TGF-β2 or 5 × 10⁵ P1 HFSCs with FITC-conjugated TGF-β2 medium or 5 × 10⁵ P1 LbL-HFSCs, combined with 5 × 10⁵ mouse dermal cells were injected subcutaneously into the dorsal site of athymic nude mice (n =16 per group). FITC- conjugated TGF-β2 were detected using the In vivo FX Pro imaging system (Bruker, Madison, WI, USA) according to manufacturer's instructions, on days 1, 3, 5, and 7 post-injection. All fluorescence intensities were analyzed using the MISE software (Bruker), and are presented in terms of photon flux (photons sec^-1, cm^-2 steradian [SR]^-1). On days 1, 3, 5, and 7 post-injection, the grafted regions were harvested and 10-µm thick frozen sections were obtained. Fluorescence microscopy images were captured via fluorescence microscopy (IX71 FL, Olympus).

Animal experiments were approved by the Eunice Kennedy Shriver NICHD animal protocol (ASP: 15-028). Six week-old female nude athymic mice (nu/nu) were purchased from The Jackson Laboratory. The weight of each mouse was approximately 20 g at experiment onset. Mice were injected into the tail vein with 1 million MTT luciferase cells in 100 μL PBS. Animals were imaged from day 7 post tail-vein injection by bioluminescence imaging. Mice injected with MTT luciferase allografts received an I.P. injection of 10 μL/g of body weight of D-luciferin (Caliper Life Sciences) dissolved in 100 μL PBS. The in vivo imaging system (Bruker) and Bruker MI SE Software were utilized to acquire and analyze the signaling. Seven days after tail vein injection, all mice were randomized into two groups, each consisting of 7 mice. Mice were either treated with solvent alone, or IDA hydrochloride intraperitoneal injection (Pfizer) with a 1 mg/kg dose every day, for seven days. Animals were re-imaged to measure the size of metastatic tumors. Euthanasia by cervical dislocation followed. Liver and spleen tumors were removed. Expression profiles of HIF-1α and HIF-2α downstream target genes were measured using qPCR according to the protocol described above.

Subcutaneous tumor volume was measured every other day with a caliper and calculated as V = (π/6) AB² (A and B = the largest and the smallest dimension of the tumor, respectively) [54 (link)]. Survival curves are based on the time of death caused by tumor growth or on the time of sacrifice of mice reaching the maximally allowed tumor size of 2 cm in diameter.
In metastatic PHEO, mice were imaged by an IVIS system (Bruker, Billerica, MA, USA) once a week to detect a bioluminescence signal intensity of metastatic organ lesions, and the signal was evaluated using Bruker MI SE software.

The animal experiment followed the Declaration of Helsinki guidelines and was approved by the Guangdong Medical University Animal Ethical Committee (GDY2102330). 5-week-old female C57BL/6 mice were obtained from Yancheng Biological Company and housed in an SPF laboratory at 26℃ with free access to water and food. Mice were intraperitoneally injected with 5 × 10⁶ ID8-Luc-puro cells in 200 μL PBS. On the seventh day after cell inoculation, mice were treated with 1 mpk trigonelline, 30 mpk ML385, 0.5 mpk clobetasol propionate, 3 mpk RSL3, 5 mpk ML210 and their respective combinations, while 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline was given as a control. Body weight was measured every 5 days. After treatment, mice were injected with pentobarbitone (1 mg/mice) and D-luciferin (3 mg/mice) successively, and luminescence signals were observed within 15 min using the In-vivo Xtreme live imaging system (Bruker, USA). Illuminance intensity was normalized and analyzed using Bruker MI SE software.

Bioluminescence imaging on live mice was previously described (25 (link), 28 (link)). Briefly, mice were anesthetized under 2% isoflurane in 100% Oz at 0.5–1 L/min., followed by intraperitoneal injection of 300 mg/kg d-luciferin (Biosynth, St. Gallen, Switzerland). Anesthetized mice were placed in a light-tight chamber of the In vivo Xtreme Imaging System (Bruker, Billerica, MA, USA), and photons emitted from live luciferase-expressing cells were captured by the back-illuminated 4 MP camera within 30 min after luciferin injection. Bioluminescent imaging (BLI) signals were expressed in photons/second by drawing a defined region of interest at the site of transplant throughout the experiment with the Bruker MI SE software.