MTT assay:
RAW Blue cell line viability was assessed by the mitochondrial
metabolization of the tetrazolium salt 3-(4,5-methyltiazol-
2yl-)-2,5diphenyl-tetrazolium bromide] (MTT) (Applichen, Germany)
colorimetric assay. Cells were maintained and plated as explained
previously. The same treatment used for QuantiBlue assay was performed.
Then MTT (0.5 mg·ml−1) was added, and cells were
incubated at 37°C for 2 hours. Cell media was removed and formazan
crystals were dissolved in DMSO. Optical density was measured at 570 nm
(OD570) on a microplate spectrophotometer (PherastarFSX,
BMG Lab, UK).
ELISA:
Measurement of the protein levels of CXCL1 and CCL2 from peritoneal
lavage fluids, or secreted into cell supernatants from 1.5 ×
106 resident peritoneal macrophages was performed by
ELISA (R&D Systems) according to the manufacturer’s instructions.