Mitoscreen jc 1 assay kit

Effect of SNG on the MMP of MM cells was determined using BD MitoScreen (JC-1) assay kit as described earlier (45 (link)). After incubation with SNG, cells in 1x assay buffer were incubated with JC-1 stain for 15 min at 37°C, washed and then analyzed by Flow Cytometry, to quantify the reduction in red fluorescence.

As described above, mouse MDSCs were induced in vitro. Then induced MDSC cells were sorted using anti-mouse Ly-6G and Ly-6C particles-DM (BD Biosciences, USA). Ly6G⁺Ly6C⁺ cells (MDSC) proliferation and cell division was assayed by using carboxyfluorescein succinimidyl ester (CFSE) (Invitrogen, USA) labeling. Cell apoptosis was assessed using a MitoScreen (JC-1) assay kit (BD Biosciences, USA) according to the manufacturer’s protocol. The labeled cells were treated with vehicle or the different concentration of L-4F (5, 10, 20 μg/ml) for 48 h. Then, the cells were assessed using BD FACSVerse flow cytometer and quantified with FlowJo 7.6 software.