Ami htx

The animal procedures in this study were approved by the Ethical Committee for Animal Research of Southern Medical University (Guangzhou, China).The BALB/c nude mice (3–4 weeks old, male) were purchased from the Central Animal Facility of the Southern Medical University. 8 × 10⁵ cells were injected into the left ventricle of the heart. Treatments for intracardiac injected mice were all started after two weeks and maintained for two weeks. The treatments were as follows: 1. Each mouse was injected with IGF1-neutralizing antibody (50 µg/mL in PBS, 5 µL/mouse) every other day. 2. Each mouse was gavaged twice daily with NVP-AEW541 (20 mg/kg). 3. Each mouse was injected intraperitoneally with rapamycin (5 mg/kg) daily. BLI data were acquired by Ami HTX (Spectral Instruments Imaging). Micro-CT data acquired by uCT80 (Bruker) system. RNA sequencing was performed to identify differentially expressed genes between 5-8F and BM3 cells. Bone tissue was collected for paraffin embedding and sectioning. HE staining was used to assess tissue condition. TRAP staining was performed with the TRAP kit to assess the number of osteoclasts.

Nine or ten weeks old female wild-type BALB/c were injected with a 2 mg Arthrogen-CIA-5-Clone monoclonal antibody Cocktail (Chondrex) through intravenous (i.v.). After 3 days, 50 μg of lipopolysaccharaide was injected intraperitoneally. On day 10, CDr17 (1 mM) was injected into CAIA-induced mice by i.v. injection. The in vivo imaging was performed by an AMI HTX (Spectral Instruments Imaging, Tucson, AZ, USA) with λ_ex = 640 nm and λ_em = 690 nm. The data were analyzed by Aura software (Spectral Instruments Imaging). The separate three experiments were proceeded with similar results. No data were excluded from the analyses.

Mice were injected intraperitoneally with 150 mg/kg D-luciferin in PBS 10 min prior to imaging. Imaging was performed using AMI HTX (Spectral Instruments Imaging, Tucson, AZ, USA). The mice were anesthetized with isoflurane prior and during the imaging (n = 9–10 mice per group). Total photon flux (photon/s) was measured from a fixed region-of-interest (ROI) over the skull using BLI and XQuartz software (version 2.7.11).

For four weeks after tumor cell inoculation, the mice were imaged every week with Ami HTX optical imaging system (Spectral Instruments Imaging). Before each imaging session, the mice were injected with D-luciferin (150 mg/Kg in saline, PerkinElmer, 122799) and exposed to a 2–5% isoflurane/oxygen mixture for anesthesia. Images were taken 8 min after injection of D-luciferin and with 20 s and 40 s exposure times.

Nine weeks old female wild-type DBA1/j mice were purchased from Central Lab Animal Inc. (Seoul, Korea). 100 µL of emulsified solution (1:1, v/v) of collagen type II (4 mg/mL) (Chondrex) and CFA (1 mg/mL) (Chondrex) mixture was intradermally (i.d.) injected into DBA/1j mice. The incidence of arthritis was complete after 5 weeks, and CDr17 (1 mM) was injected into CIA mice by i.v. injection. The in vivo imaging was performed by an AMI HTX (Spectral Instruments Imaging, Tucson, AZ, USA) with λ_ex = 640 nm and λ_em = 690 nm. The data were analyzed by Aura software (Spectral Instruments Imaging). The separate three experiments were proceeded with similar results. No data were excluded from the analyses.