Anti mouse cd86 bv785

Female C57BL/6 mice (6–8 weeks) were purchased from Shanghai SLAC Co. Ltd. All in vivo experiments were performed according to protocols approved by the Laboratory Animal Center of The Second affiliated Hospital of Zhejiang University School of Medicine. Mice were randomly divided into six groups (n = 5), including: (1) PBS, (2) PBS + US (3) PPIX NDs, (4) PPIX ND + US, (5) PMPS NDs and (6) PMPS NDs + US. PanC02 cells ( ${2\times 10}^6$ ) were orthotopically injected into the pancreas of each mouse. Seven days later, the tumors were allowed to reach 50 ~ 100 mm³ before the experiments. The mice in other groups were administered with PBS, PPIX NDs (PPIX 10 mg/kg), or PMPS NDs (equal dose of PPIX:10 mg/kg). In US irradiation group, US (Frequency: 3 MHz; Acoustic intensity: 1.0 W/cm²; Duty cycle: 20%; Times: 5 min) was conducted at five minutes and 6 h post-injection. On the third day, the tumor-draining lymph nodes were excised for analysis by FCM after co-staining with anti-mouse CD11c BV421 (Biolegend, 117329), anti-mouse CD80 PE/cyanine7 (Biolegend, 104733), and anti-mouse CD86 BV785 (Biolegend, 105043). Meanwhile, the blood samples were collected at 72 h after treatments, and the proinflammatory cytokines, including IL-6 and TNF-α, were tested by ELISA.

According to an established method, Bone-marrow-derived DCs (BMDCs) were isolated from 6–8-week C57BL/6 mice. The PanC02 cells after different treatments (PBS, PBS + US, PPIX NDs, PPIX NDs + US, PMPS NDs, PMPS NDs + US) were incubated with BMDCs respectively. After 2-day stimulation, the BMDCs were DCs stained with anti-mouse CD11c BV421 (Biolegend, 117329), anti-mouse CD80 PE/cyanine7 (Biolegend, 104733), and anti-mouse CD86 BV785 (Biolegend, 105043) were analyzed by FCM (Beckman CytoFlex).