Fig. 2

Astrocytes exhibit temporal expression of autophagy-related genes and autophagy flux in response to Aβ oligomer in a timely manner. a, RNA-seq analysis in human astrocyte culture at 1, 3, 6, 9 and 12 h after Aβ oligomer treatment. The altered gene expression was categorized into four groups: Group I, II, III and IV depending on the time point of peak expression. b, Experimental design for a time course of qRT-PCR in Aβ oligomer-treated human astrocytes. c, Heatmap of the expression pattern for autophagy-associated genes. d, qRT-PCR analysis for LC3B, SQSTM1 and BECN1 mRNA levels in response to Aβ oligomer. Line graphs present the mean ± SEM of three separate experiments. e, Western blot analysis data showing that levels of endogenous LC3B-II and SQSTM1 are time-dependently changed in Aβ oligomer-treated primary mouse astrocytes. f, Quantification of the band intensities of LC3B-II and SQSTM1 normalized by β-actin (ACTB). Line graphs present the mean ± SEM of three separate experiments. g, Double immunostaining with LC3B and SQSTM1 antibodies in monomeric or oligomeric Aβ-treated human astrocyte culture. Scale bars (white): 20 μm. h & i, Bar graphs showing the average intensity of LC3B (h) and SQSTM1 signals (i). A total of 20 cell count, 5 cells/well, n = 4 wells. Data are presented as mean ± SEM. Significantly different at *, p < 0.05; **, p < 0.01