In parallel, the effects of • NO scavenging on cell migration were evaluated via transwell migration and wound healing assays. For the transwell assay, after overnight incubation of cells in FBS-free RPMI, they were trypsinized, counted and 80,000 cells were seeded onto the upper chamber of each transwell insert in FBS-free RPMI, and placed in a well of a 24-multiwell plate. The cell migration conditions were optimized, and the final study involved diluting DETA-NO in the FBS-containing medium, which were placed in the well to a final concentration of 300 μM with/without different concentrations of the tested compounds. After 24 hours of culturing, the cells were fixed in PFA, washed and the non-migrated cells were cleaned with a cotton swab. The migrated cells in the lower surface of the membrane were stained with 1 μg/mL DAPI. After washing with PBS, these cells were imaged using an Inverted Fluorescence microscope and counted using ImageJ software. For the wound healing assay, 50,000 cells were added to each well of a sterile 2 well silicone insert placed in a 24-multiwell plate, and left to attach for 24 hours. Next, the inserts were removed and the cells were washed with PBS and treated with 300 μM DETA-NO diluted in phenol red-containing and FBS-free RPMI with/without different concentrations of the tested compounds. Each well was then imaged each per hour using the incucyte system for a period of 24 hours. The analysis of the wound healing rate used ImageJ software and MiToBo toolbox (Möller et al., 2016). Three samples were tested per group, the experiment was performed two times, and the averages of all measurements were taken.