Filipin 3

After treatment, the cells were collected and washed by PBS for 2 times and smear on the cover glass, and then fixed with 3% paraformaldehyde for 20 min, washed by PBS and incubated with 1.5 mg/mL glycine for 10 min. The cells were rinsed with PBS and incubated with 50 μg/mL Filipin III (B6034, APExBIO Technology, TX, USA) containing 10% FBS for 2 h at room temperature in the dark, followed by observed with a confocal laser scanning microscope.

Ovalbumin (OVA) was purchased from Sigma-Aldrich (USA). CpG oligonucleotide 1826 (5ʹ-TCC ATG ACG TTC CTG ACG TT-3ʹ) was synthesized by Sangon (China). Fetal bovine serum (FBS), phosphate-buffered saline (PBS), RPMI-1640, DMEM were purchased from Hyclone (USA). Rottlerin, chlorpromazine, Filipin III and cytochalasin D were purchased from ApexBio Technology (USA). Anhydrous dimethyl sulfoxide (DMSO), red blood cell lysis, the carbocyanine dye Dil, 4ʹ,6-Diamidino-2-phenylindole dihydrochloride (DAPI, ≥90%), fluorescein isothiocyanate (FITC) and near-Infrared Cyanine 7 dyes (Cy7 NHS ester) were purchased from Solarbio Science & Technology Co. Ltd (China). Cell Counting Kit-8 (CCK-8), NP-40 lysis buffer, X-Gal (ST912) and BCA protein detection kit were purchased from Beyotime (China). Recombinant mouse IL-4 and GM-CSF were purchased from Peprotech (USA). Anti-mouse monoclonal antibodies (CD80 (B7-1), CD86 (B7-2), CCR7, CD40, MHC I (H-2Kb) and MHC II (I-Ab)) were purchased from Biolegend (USA). None of the chemicals were further purified.