I a 1 e mhc 2 pe cy7 m5 114

The effects of SFSHs on the maturation and activation of dendritic cells were investigated in vitro. DC2.4 cells were cultured in RPMI 1640 medium supplemented with 10% (v/v) fetal bovine serum (FBS) and 1% (v/v) antibiotics (penicillin-streptomycin, PS) in a cell incubator (37 °C, 5% CO₂). Cells were seeded in 24-well plates at a density of 10⁵ cells/well, following the treatment of PBS (10 μl), free OMVs (10 μl), regular polyacrylamide hydrogels (10 μl), or SFSHs (10 μl) for 20 h and 40 h, respectively. Then, cells were collected and washed with PBS. For specific labeling, cells were stained with anti-mouse antibody against CD86-APC (GL-1, Cat: 105012, BioLegend), CD80-PE (16-10A1, Cat: 104708, BioLegend), I-A/I-E (MHC II)-PE/Cy7 (M5/114.15.2, Cat: 107629, BioLegend) in 0.5% bovine serum albumin (BSA) dissolved in PBS on ice for 1 h. Antibodies were used at a dilution of 1:200. After washing with PBS for 3 times, cells were analyzed by FCM.