Fig. 5

AS-IV promotes mitophagy and decreases the mitochondrial ROS production in astrocytes. Astrocytes were cultured for 10 days or 40 days in vitro (DIV) and then treated with AS-IV (50 μM) for 10 days. a–f Representative immunoblots (a) and quantitative analysis of p62 (b) and LC3 (c) in the cytoplasm and PINK1 (d), Parkin (e), and TOM20 (f) in the mitochondria of astrocytes. Quantified data are normalized to the control group (the control group value is equal to 1). g The scatter plot of the flow cytometry analysis shows the distribution of gating of JC-1 red-positive (aggregates) and JC-1 green-positive (monomers) cell population. h The bar graph shows the percentage of JC-1 monomer-positive cells. i Astrocytes was stained with MitoSOX and analyzed by flow cytometry. Representative dot plots are shown illustrating the gating of MitoSOX-positive (R2) populations. The percentages of MitoSOX-positive (R2) populations represent relative mitochondrial ROS levels. j Quantification of the mitochondrial ROS. The data shown are the mean ± SEM from three independent experiments. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001. AS-IV, Astragaloside IV