Physiological properties of astrocyte subtypes. (A) ESC-derived astrocyte RNA-seq expression of calcium signaling- and membrane polarization-related genes in astrocyte subtypes (n=3 biological replicates, error bars represent s.d. of TPM). (B,C) I-V curves for subtype astrocytes measuring inward whole-cell current (B) and membrane resistance of astrocyte subtypes (C) (n=3 biological replicates, six cells measured per replicate, 18 total per region patched; *P<0.05, Student's t-test). (D) Average response (black line) and s.d. (colored bars) of ATP-induced calcium release for astrocyte subtypes visualized with Fluo-4AM (n=3 biological replicates totaling >50 analyzed cells per region). (E) Examples of characteristic responses of astrocytes to ATP visualized with Fluo-4AM. The four characteristic intracellular calcium release responses to ATP identified in all regional astrocyte populations are shown. (F) Percentage of cells exhibiting the calcium oscillations shown in panel E (n=3 biological replicates totaling >50 cells analyzed, error bars represent s.d.). (G) Maximum change in fluorescent intensity in the presence or absence of 2-APB (50 μM), dantrolene (10 μM) or ryanodine (50 nM) after the addition of ATP (n=3 biological replicates totaling >50 cells analyzed, error bars represent s.d.; *P<0.01, Student's t-test). n.s., not significant.