Ti2 e a1

Immunofluorescence colocalization experiments were performed on three thyroid cancer specimens to observe the positional relationship between IL‐6 and M2‐phenotype macrophages within the tumor. Briefly, the tumor tissue sections were incubated with primary antibodies against IL‐6 (21865‐1‐AP, Proteintech Group Inc) and CD206 (18704‐1‐AP, Proteintech Group Inc), a specific marker of the M2 phenotype, and then incubated with Alexa Fluor 492–labelled donkey anti‐rabbit IgG(H + L) and Alexa Fluor 550–labelled donkey anti‐rabbit IgG(H + L). Nuclei were counterstained with DAPI. Fluorescent images were obtained by a confocal laser‐scanning microscope (Ti2‐E‐A1, Nikon).

The PNPs were loaded onto the surface of ASEc through electrostatic interactions. PNPs were added and co-incubated with ASEc for 30 min. The ASEc@PNPs complexes were collected by centrifugation (4°C, 5000 rpm, 5 min) and washed three times with sterile PBS.
The viability of nanoparticle-loaded ASEc was assessed using the plate colony-counting method. Briefly, ASEc@PNPs complexes were diluted with sterile PBS at 100:1 and coated onto LB agar plates, after which the plates were cultured at 37°C overnight. The number of colonies on the plates was counted to calculate the viability of PNPs-loaded ASEc.
The morphology of the ASEc@PNPs complexes was investigated using TEM. DLS measured the complex’s size distribution and zeta potential. The ASEc@PNPs pellet was collected and stained with DAPI for 20–30 min. Confocal fluorescence images (CLSM, Ti2-E+A1, Nikon, Japan) were used to observe the cell morphology.