Rs 2000 pro x ray bio irradiator

This study was approved by the Institutional Animal Care and Use Committee of Nanjing Medical University (IACUC-1901012). BALB/c nude mice were purchased from the Animal Center of Nanjing Medical University. A total of 2 × 10 6 cells were injected subcutaneously into the anks of nude mice.
Tumours were measured by callipers every 5 days using the following formula: volume = (width 2 × length)/2. When the tumour volume reached 50 mm 3 , 10 µg (50 µl) EVs or 50 µl PBS were injected into tumour once daily for 5 days. Meanwhile, tumours were irradiated by an RS 2000 Pro X-Ray Bio-irradiator (Radsource, USA) with 2 Gy per day for 4 consecutive days starting from the second day of EV injection. Two weeks post-IR, the mice were euthanized. In addition, we injected NC-Te13 cells and miR-340-5p mimics Te13 cells into the nude mouse anks to study the effect of miR-340-5p. When the tumour volume reached 50 mm 3 , four fractions of radiation (2 Gy/f) were applied to the tumours, and the mice were sacri ced 14 days after the radiation was completed. Lead shields were used to prevent radiation injury.

Human OSCC cell lines (Te13, Te1 and Eca109) were obtained from American Type Culture Collection (ATCC, USA). All cell lines were cultured in RPMI-1640 medium (Gibco, USA) with 10% foetal bovine serum (FBS; Gibco, USA), 100 U/ ml penicillin and 100 µg/ml streptomycin. Cells were maintained at 37 °C and 5% CO 2 and were routinely examined for Mycoplasma contamination. To induce hypoxia (< 1% O 2 ), cells were cultured at 37℃ in the same incubator in an AnaeroPack jar with AneroPack-Anaero (Mitsubishi, Japan) according to the manufacturer's instructions. The hypoxic environment was con rmed by hypoxia inducible factor 1 subunit alpha (HIF-1α) expression. Cells were irradiated by an RS 2000 Pro X-Ray Bioirradiator (Radsource, USA).

This study was approved by the Institutional Animal Care and Use Committee of Nanjing Medical University (IACUC-1901012). BALB/c nude mice were purchased from the Animal Center of Nanjing Medical University. A total of 2×10 6 cells were injected subcutaneously into the flanks of the nude mice. Tumours were measured with calipers every 5 days, and the tumour volume was calculated using the following formula: volume = (width 2 × length)/2. When the tumour volume reached 50 mm 3 , 10 µg (50 µl) of EVs or 50 µl of PBS was injected into the tumour once daily for 5 days. In addition, tumours were irradiated in an RS 2000 Pro X-Ray Bio-irradiator (Radsource, USA) with a dosage of 2 Gy per day for 4 consecutive days starting on the second day of EV injection. Two weeks post IR, the mice were euthanized. In addition, we injected NC-Te13 cells and miR-340-5p mimic Te13 cells into the flanks of nude mice to study the effect of miR-340-5p. When the tumour volumes reached 50 mm 3 , four fractions of radiation (2 Gy/f) were applied to the tumours, and the mice were sacrificed 14 days after the radiation exposure was completed. Lead shields were used to prevent radiation injury.

Human OSCC cell lines (Te13, Te1 and Eca109) were obtained from the American Type Culture Collection (ATCC, USA). All cell lines were cultured in RPMI-1640 medium (Gibco, USA) with 10% foetal bovine serum (FBS; Gibco, USA), 100 U/ml penicillin and 100 μg/ml streptomycin. Cells were maintained at 37°C in 5% CO 2 and were routinely examined for Mycoplasma contamination. To induce hypoxia (<1% O 2 ), cells were cultured at 37°C in the same incubator in an AnaeroPack jar with AnaeroPack-Anaero (Mitsubishi, Japan) according to the manufacturer's instructions. The hypoxic environment was confirmed by detection of hypoxia inducible factor 1 subunit alpha (HIF-1α) expression. Cells were irradiated by RS 2000 Pro X-Ray Bio-irradiator (Radsource, USA) with 140kV X-ray beam. The irradiation field was confined within the culture dish or disk, and the dosing rate was 1.439Gy/min.