Extended Data Fig. 2: Validation of iAstrocyte differentiation from two additional hiPSC lines.

A, Representative images of immunofluorescence against GFAP, S100β, GLAST, Cx43, glutamine synthetase, or vimentin in iAstrocytes vs. TCW astrocytes derived from TCW-1E44 or 162D hiPSCs (scale bar: 60 μm). b,Quantification of GFAP, S100β, GLAST, Cx43, glutamine synthetase, or vimentin immunofluorescence intensity (n = 3 wells). c, Phagocytosis of pHrodo-labeled rat synaptosomes (median fluorescence intensity measured by flow cytometry) by iAstrocytes derived from TCW-1E44 or 162D hiPSCs in the absence (n = 5 wells) or presence (n = 1 well) of cytochalasin D (cytoD). d, Percent VCAM1+ cells in TCW-1E44 or 162D iAstrocytes treated with vehicle control vs. IL-1α+TNF+ C1q (n = 4 wells). e, Percentage of dead cells (measured by TO-PRO-3 permeability) in iNeurons incubated with conditioned media from TCW-1E44 or 162D iAstrocytes treated with vehicle control or IL-1α+TNF+ C1q (n = 12 wells). In panels b and c, P values were calculated using the two-sided Student’s t-test. In panels d and e, P values were calculated using the two-sided Mann-Whitney U test.