Fig. 5

Effect of Drd2 activation on the interaction of β-arrestin2 and NLRP3. A Heatmap of 4D label-free mass spectrometry-quantified proteins that bound β-arrestin2 (n = 2). B Representative fragmentation spectrum of FLQESNVLYQHNLR in STAT3. Flag-tagged STAT3 construct and β-arrestin2 pcDNA3.1 construct was co-transfected in HEK293T cells. C Immunofluorescent staining for β-arrestin2, DAPI, and STAT3 in HEK293T cells. Green: β-arrestin2; Red: STAT3; Blue: DAPI. Scale bar: 20 μm. D Cell lysates were immunoprecipitated with anti-Flag antibody and then the samples were analyzed by immunoblotting. E Cell lysates of primary astrocytes treated with IL-6 (300 ng/mL) and UNC9995 or not were immunoprecipitated with anti-STAT3 antibody, and then the samples were analyzed by immunoblotting. F Cell lysates of primary astrocytes treated with IL-6 (300 ng/mL) and UNC9995 or not were immunoprecipitated with anti-β-arrestin2 antibody, and then the samples were analyzed by immunoblotting. G Flag-tagged STAT3 construct was transfected in primary astrocytes. Cell lysates of primary astrocytes treated with IL-6 (300 ng/mL) and UNC9995 or not were immunoprecipitated with anti-flag antibody, and then the samples were analyzed by immunoblotting. H Primary astrocytes were pretreated with UNC9995 for 1 h and then stimulated with IL-6 (300 ng/mL) for 24 h. β-Arrestin2 and STAT3 proximity ligation signals. Green: FITC–phalloidin; Red: STAT3–β-arrestin2; Blue: DAPI. Scale bar: 20 μm