Crocodile 4 in one assay miniworkstation

ELISA was performed to measure the protein expression levels of cellular cytokines (IFN-γ, IL-17A and IL-22) in the peripheral blood of patients with VIT and of healthy individuals, or from CD4⁺T cells or melanocyte supernatant. The PBMCs and supernatant were separated by centrifugation at 2,000 × g for 15 min at 4°C and stored in aliquots at 80°C. The expression levels of inflammatory factors (IFN-γ, IL-17A and IL-22) in CD4⁺T cells or melanocyte supernatant were determined using their corresponding ELISA kits (cat. nos. KHC4021 for IFN-γ, BMS2017 for IL-17A and BMS2047 for IL-22; Invitrogen; Thermo Fisher Scientific, Inc.). OD values were detected and measured using a Crocodile 4-in-one assay miniWorkstation (Titertek-Berthold).

Levels of inflammatory cytokines TNF-α (catalog no. BMS223-4), Interleukin-6 (IL-6, catalog no. EH2IL6) and Interleukin-8 (IL-8, catalog no. KAC1301) in HPAEC supernatant were quantified with their corresponding ELISA kits (Invitrogen) following the manufacturers’ manuals. OD values were measured using a Crocodile 4-in-one assay miniWorkstation (11050011, Titertek-Berthold, Pforzheim, Germany).