Fig. 5

β-arrestin1 co-localizes with Drp1 in primary astrocytes. A Identification of Drp1 as a β-arrestin1-associated protein. Mass spectrometry spectrum of a Drp1 tryptic peptide. B Flag-tagged Drp1 construct and β-arrestin1 pcDNA3.1 construct were co-transfected in HEK293T cells. Representative immunofluoresent staining of β-arrestin1 (green) and Drp1 (red) processed by Imaris Microscopy Image Analysis software. C Flag-tagged Drp1 construct and β-arrestin1 pcDNA3.1 construct were co-transfected in HEK293T cells. Cell lysates were immunoprecipitated with anti-Drp1 antibody and then the samples were analyzed by immunoblotting. D Flag-tagged Drp1 construct and β-arrestin1 pcDNA3.1 construct were co-transfected in HEK293T cells. Cell lysates were immunoprecipitated with anti-β-arrestin1 antibody and then the samples were analyzed by immunoblotting. E Immunoblotting analysis of Fis1 proteins in cell lysates of primary astrocytes immunoprecipitated with Drp1 antibody. F Immunoblotting analysis of Drp1 proteins in cell lysates of primary astrocytes immunoprecipitated with Fis1 antibody. G Protein levels of Drp1 in cytoplasmic and mitochondrial parts. H Immunofluorescent staining of TOMM20 (green) and Drp1 (red) in primary astrocytes. I Relative co-localized signals of the TOMM20-positive and Drp1-positive immunofluorescent particles between groups. J Representative images of Mitotracker Green in primary astrocytes under confocal microscope. Data were analyzed by two-way ANOVA followed by Tukey's multiple comparisons test. ^*P < 0.05 and ^***P < 0.001 vs. the WT CON-MCM group. ^##P < 0.01 vs. the WT LPS-MCM group. Values are presented as means ± SEM from three independent experiments