Cell total iron colorimetric assay kit

Total iron and Fe²⁺ contents in trophoblast cells were measured using a Cell Total Iron Colorimetric Assay Kit (E-BC-K880-M; Elabscience, Wuhan, China) and Cell Ferrous Iron Colorimetric Assay Kit (E-BC-K881-M; Elabscience), respectively, and a ReadMax 1900 microplate reader (Shanghai Flash, Shanghai, China) was used to measure absorbance at 593 nm. GSH and GSSG concentrations were detected using a GSH and GSSG Assay Kit (S0053; Beyotime) and by measuring the cell absorbance at 412 nm.

Pg strain ATCC33277 was obtained from BeNa Culture Collection Center (Beijing, China). Anti‐CD63 & anti‐TSG101 were bought from Umibio (Shanghai, China). Exosome‐depleted FBS was provided by HAKATA (Shanghai, China). Exo‐spin mini standard and exo‐spin buffer were bought from TheWell Bioscience (North Brunswick, USA). LPS, hemin, and transferrin were purchased from Sigma‐Aldrich (St. Louis, USA). Mouse IL‐4 was purchased from PeproTech (RockyHill, USA). Firefly luciferase reporter gene lentiviral vector was bought from Genechem (Shanghai, China). DiR and FITC were bought from Beyotime Biotechnology (Shanghai, China). DCFH‐DA and Rosup were bought from Solarbio (Beijing, China). HO‐1 Assay Kit, Ferrous Iron Colorimetric Assay Kit, and Cell Total Iron Colorimetric Assay Kit were provided by Elabscience (Wuhan, China). n situ Apoptosis Detection Kit was bought from TaKaRa (Beijing, China). APC‐CD11b, FITC‐CD206, Percp/Cy5.5‐CD80, PE‐CD86, FITC‐CD3, PE‐CD4, Percp/Cy5.5‐CD8, FITC‐PD1 was bought from BioLegend (San Diego, USA). Mouse IL‐4 and Mouse IL‐1 ELISA Kits were bought from Boster (Pleasanton, USA).

The Cell Total Iron Colorimetric Assay Kit was purchased from Elabscience (Wuhan, CN) to conduct Fe²⁺ level assay. The transfected cells were lysed on ice and centrifuged at 15000 g. We then collected the supernatant and added it to a reagent containing iron reductase. After incubating for 40 min, the optical density was measured by a microplate reader at 593 nm.