Figure 1.

Induction of a reactive phenotype in hPSC-derived astrocytes

(A and B) Reactive astrocytes exhibited a hypertrophic profile as seen by S100β and GFAP staining.

(C and D) Increased accumulation of C3 staining in the perinuclear region of reactive astrocytes.

(E–H) Morphological analysis showed an increased number of branches and shorter processes in reactive astrocytes, as well as an overall increased number of process intersections through Sholl analysis.

(I and J) Increased C3 expression exhibited by reactive astrocytes through western blot analysis.

(K) qPCR analysis showed that reactive astrocytes express increased levels of pan-reactive associated genes, as well as genes that specifically characterize the phenotype of neurotoxic reactive astrocytes. Results are represented as fold change vs. control astrocytes in (J) and (K). Colored dots correspond to 3 different lines, H7 hPSC line in blue, mips2 hPSC line in gray, and WTC11 hPSC line in orange. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 vs. control samples, two-tailed paired Student’s t test. Scale bar: 30 μm in (A) and (B), 20 μm in (C) and (D). Data represents mean values ± SEM.