Fig. 6

Effects of HSF1 agonist 17-AAG on the conversion of astrocyte phenotypes. a Toxic assay of A1-astrocyte-conditioned medium (ACM) containing 3 ng/ml IL-1α, 30 ng/ml TNF-α, and 400 ng/ml C1q in the DMEM for the astrocytes. b–d Effects of ACM on the conversion of astrocytes evaluated by C3 (b, c) and S100A10 (b, d) protein levels following cell incubation for 24 h. Quantities were normalized to endogenous β-actin. Experiments were performed in triplicates. Error bars represent the standard deviation, *P < 0.05. The data were analyzed by two­tailed unpaired Student’s t test. P = 0.0002 in (c) and P = 0.0042 in (d). e–i Determination of C3 (e, g), S100A10 (e, h), and HSP70 (e, i) in astrocytes following cell incubation at ACM for 24 h in the presence of 0-10 μmol/L 17-AAG. f Toxic assay of ACM in the presence of 0–10 μmol/L 17-AAG for the astrocytes. Quantities were normalized to endogenous β-actin. Experiments were performed in triplicates. Error bars represent the standard deviation, *P < 0.05, ^#P < 0.05, one-way ANOVA with Tukey’s post hoc test. For C3 quantitative analysis in (g), F (7, 16) = 66.95, P = 0.0207. ACM vs con, P < 0.0001; 10 μmol/L 17-AAG + DMSO vs ACM, P < 0.0001; 0.1 μmol/L 17-AAG + ACM + DMSO vs ACM, P = 0.0103; 0.5 μmol/L 17-AAG + ACM + DMSO vs ACM, P < 0.0001; 2.5 μmol/L 17-AAG + ACM + DMSO vs ACM, P < 0.0001; 10 μmol/L 17-AAG + ACM + DMSO vs ACM, P < 0.0001. For S100A10 quantitative analysis in (h), F (7, 16) = 4.324, P = 0.0073. ACM vs con, P = 0.018. For HSP70 quantitative analysis in (i), F (7, 16) = 19.79, P < 0.0001. ACM vs con, P = 0.8082; 10 μmol/L 17-AAG + DMSO vs ACM, P = 0.0019; 0.1 μmol/L 17-AAG + ACM + DMSO vs ACM, P = 0.0029; 0.5 μmol/L 17-AAG + ACM + DMSO vs ACM, P < 0.0001; 2.5 μmol/L 17-AAG + ACM + DMSO vs ACM, P = 0.0015; 10 μmol/L 17-AAG + ACM + DMSO vs ACM, P < 0.0001