Rhodamine phalloidin

Ferric acetylacetonate (AR, Sinopharm Chemical Reagent Co. Ltd., SCRC, Shanghai, China), oleic acid (85%, Aladdin, Shanghai, China), benzyl ether (98%, Sigma-Aldrich, St. Louis, MS, USA), ethanol (AR, SCRC), n-hexane (97%, SCRC), dimercaptosuccinic acid (98%, TCI, Shanghai, China), acetone (AR, SCRC), triethylamine (AR, Aladdin, Shanghai, China), methanol (AR, SCRC), polyethyleneimine (Mw 25000, Sigma-Aldrich, St. Louis, MS, USA), heptafluorobutyric anhydride (97%, Macklin, Shanghai, China), amino-polyethylene glycol-carboxyl (Mw 2000, Xi’an Ruixi, Xi’an, China), methoxy-polyethylene glycol-polyethyleneimine (mPEG-PEI, Mw 2000-25000, Xi’an Ruixi Co. Ltd., Xi’an, China) FITC (Beyotime, Shanghai, China), CCK8 kit (KeyGen, Nanjing, China), Hoechst33342 (KeyGen, Nanjing, China), rhodamine-phalloidin (KeyGen, Nanjing, China), LysoTracker (KeyGen, Nanjing, China), Annexin V-FITC/PI kit (KeyGen, Nanjing, China), Anti-CXCR4 antibody (Abcam of Thermo Fisher Scientific Inc., Waltham, MA, USA), DMEM high glucose medium (KeyGen, Nanjing, China), fetal bovine serum (Gibco of Thermo Fisher Scientific Inc., Waltham, MA, USA), trypsin (Gibco of Thermo Fisher Scientific Inc., Waltham, MA, USA), siRNA(GenePharma Co. Ltd., Shanghai, China), 4 T1, A549, HeLa cell line was provided by Southeast University School of Medicine (Nanjing, China).

The animal tissues were prepared as described above including fixation, decalcification, tissue embedding, and section. After deparaffinization and rehydration, samples were blocked with serum. RAW 264.7 cells were fixed in 4% paraformaldehyde. After permeabilization and blocking in 5% nonfat dry milk in PBS, the tissue sections or cultured cells were incubated with anti‐IDO antibody (Cell Signaling Technology) or anti‐AhR antibody (Proteintech). Next, tissue sections or cells were incubated with anti‐IgG antibody conjugated with Alexa Fluor 488 (Invitrogen). Rhodamine‐phalloidin (KeyGen) was used for stain actin filaments. 4′,6‐diamidino‐2‐phenylindole (DAPI) (Beyotime) was used to stain the nuclei. Pictures of immunofluorescent staining were visualized with confocal laser microscopy (LSM980, Zeiss).