Fig. 3
P2Y6R inhibition or knockout prevents LPS-induced phagocytosis and neuronal loss in culture. a Bright field and fluorescent images of differentiated TAMRA-stained PC12 (red), co-cultured for 3 days with IB4-labelled BV-2 (green), nuclei stained with Hoechst 33342 (blue) ± LPS. BV-2 phagocytosis of PC12 is visible as punctate TAMRA staining in IB4-stained cells (white arrowheads). Representative images; scale bar = 50 µm. b Quantification of BV-2 phagocytosis of PC12 cells ± LPS ± MRS2578 after 3 days. c Mixed neuronal–glial cultures from cerebella of wild-type (WT) or P2ry6−/− mice were treated for 3 days with ± 500 ng/ml LPS, then cell viability was measured by MTT assay. Data represent mean ± SEM of at least three independent experiments. Statistical analysis was performed using one-way ANOVA with post hoc Tukey’s multiple comparison test. ***p < 0.001 compared to vehicle-treated control (first bar). +++p < 0.001 compared to LPS-treated vehicle control (second bar). #p < 0.05 compared to LPS-treated wild-type condition (second bar)