Fig. 3

UDP/P2Y6R signaling involved in the activation of microglia cells and the production of inflammatory cytokines. a The standard curve and the UDP concentration in the LPS-treated and control supernatants of BV-2 cells based on HPLC. b mRNA levels of MIP-2 in LPS, LPS+Apy- and LPS+MRS-treated BV-2 cells. c ELISA of MIP-2 secretion. d Morphological changes in LPS-stimulated and Apy/MRS pretreated primary microglia cells. Representative immunostained images for primary microglia cells (Iba-1, red; DAPI, blue). Scale bar = 100 μm. e Representative RT-PCR showing the expression levels of inflammatory cytokines (TNF-α, iNOS, IL-6, COX-2, and MIP-2) from LPS-stimulated primary microglia cells pretreated with Apy/MRS. The band intensity was quantified using studio lite imager and is presented relative to the level of β-actin. Data are shown as the mean ± SD of three independent experiments. *p < 0.05, ***p < 0.001 vs Con, ^# p < 0.05, ^## p < 0.01, ^### p < 0.001 vs LPS. Apy (Apyrase, nucleotide hydrolase), MRS (MRS2578, P2Y6 inhibitor)