Cytokine and antibody arrays
Human Inflammation Antibody Array C3 (RayBiotech) was used to detect cytokines and factors in conditioned media from cells under indicated conditions. Cells were plated in 6-well and treated with IDX, cis or combination of both. 72 h later, culture medium was centrifuged prior to assay on the human cytokine array according to the manufacturer’s guidelines.

Western blot

Cells were lysed in RIPA buffer containing protease and phosphatase inhibitors on ice for 45 min and collected by centrifuging at 150,000 rpm for 15 min at 4 oC. Protein concentrations were measured by the bicinchoninic acid protein assay kit (Pierce). Equal amounts of protein were solubilized in laemmili buffer, boiled for 5 min and then separated by 10 % SDS-PAGE before transferring them onto nitrocellulose membranes. Membranes were blocked and probed with specific primary antibodies (Supplementary Table 2), detected by using appropriate HRP-conjugated secondary antibodies and visualized by incubation with enhanced chemiluminescence reagent (ECL, GE Healthcare) and exposure to film.