Fig. 6

Pathway analysis revealed altered status of cell-adhesion, mTOR, and cytokine release in Alexander disease astrocytes. a Schema of pathway analysis and upstream prediction analysis. b Whole-cell lysates of iPSC-derived astrocytes were prepared and equivalent amounts of total protein were loaded per lane on a polyacrylamide gel for Western blot (WB) analysis using N-cadherin or GAPDH antibodies shown on the left. c Astrocytes were immunostained with an antibody against N-cadherin (green color) and F-actin (red color) was also visualized by using Rhodamine phalloidin. Scale bar = 10 μm. d Whole-cell lysates were prepared and equivalent amounts of total protein were loaded per lane on a polyacrylamide gel for WB analysis using the panel of antibodies shown on the left. The values shown below each blot represent the ratio of phosphorylated/total band densities, calculated and normalized to the ratio in “HC1” astrocytes. e Quantification of cytokine release from iPSC-derived astrocytes. Gray/pink-colored columns indicate astrocytes of healthy controls/Alexander disease. Black-colored column indicates healthy astrocytes with addition of LPS as positive control of cytokine release. LPS: lipopolysaccharide. (*, p < 0.05, N.S.: not significant) Data represent mean ± SD (biological replicates, n = 3)