Evos fl auto 2 confocal microscope

Donor BMSC‐derived EVs were incubated with 1 µM PKH67 linker (Shanghai Umibio Co., Ltd., China) for 5 min and centrifuged. Next, imDCs were treated with PKH67‐labeled sEVs for an additional 24 h. The uptake of sEVs by imDCs was visualized using an EVOS™ FL Auto 2 confocal microscope (Thermo Fisher).

Frozen sections of graft kidneys were thawed at room temperature and washed with PBS. After this, sections were incubated with a blocking solution and then incubated with primary antibodies overnight at 4℃. On the second day, sections were incubated with a secondary antibody and then counterstained with DAPI. An EVOS™ FL Auto 2 confocal microscope (Thermo Fisher) was used to visualize the stained sections. The primary antibodies used in this experiment were as follows: integrin αE antibody (OX62) PE (Santa Cruz Biotechnology, USA), HLA‐DRA polyclonal antibody (Proteintech, USA), and CD86 polyclonal antibody (Proteintech). The secondary antibody used in this experiment was purchased from Boster Biological Technology Co., Ltd. (China).