Nicotinamide nucleotides were assayed using the NADP/NADPH assay kit according to the manufacturer’s instructions. Primary microglia were seeded onto PDL-coated 12-well plates and treated with the given concentrations of LPA for the indicated time periods. NADP and NADPH were extracted using the ... read more

The accumulated total nitrate levels were measured in the supernatant of microglia cells that were incubated with LPS in the absence or presence of an autotaxin antagonist (PF8380). This assay is based on the enzymatic conversion of nitrate to nitrite by the enzyme nitrate reductase, followed by the... read more

Intracellular levels of reactive oxygen species (ROS) of control and LPA 1 µM treated cells for the indicated time periods were quantified using the ROS-ID® Total ROS Detection kit. The kit is easy and fast to use. Thirty minutes before the end of each treatment, the cells are loaded with the ROS ... read more

We sought to determine the range of non-toxic concentrations for the pharmacological inhibitors PF8380 before being used throughout our study. Incubation of BV-2 cells with increasing concentrations of PF8380 indicated a decrease in MTT reduction, by 70% at the highest concentration used (30 µM, 24 ... read more