Figure 4.

Sox9 and Nfib improve conversion of human embryonic fibroblasts to induced astrocytes

(A) Representative immunofluorescence images of HEFs 5 weeks after induction of Sox9, Nfia, and Nfib (SAB) or Sox9 and Nfib (SB).

(B) Quantification of yield and purity of S100B, GFAP, and S100B/GFAP immunoreactive cells at 3 weeks.

(C–E) Representative maximum-intensity projection immunofluorescence images of glutamate transporters and GS in SB-HEF-iAs at 5 weeks.

(F) Glutamate uptake in SAB-HEF-iAs and SB-HEF-iAs at 5 weeks.

(G and H) Representative maximum-intensity projection immunofluorescence images of KIR4.1 and ATP1B2 in SB-HEF-iAs at 5 weeks.

(I) Example image of whole-cell patch-clamp recording of SB-HEF-iAs with patch pipette indicating a recorded cell.

(J and K) Resting membrane potential (RMP) and input resistance (Ri) values.

(L) Values of maximum current induced by ramp depolarization protocol.

(M) Current trace examples induced by ramp depolarization.

(N) Examples of current traces induced by voltage steps ranging from −160 to +20 mV. Dotted lines indicate portion of the trace where steady-state currents were quantified.

(O) Current/voltage (I/V) curve typical for astrocytes built from data obtained in (N).

hAs, primary human fetal astrocytes. Individual color levels are adjusted for visibility in (A). Data are presented as mean ± SEM of n = 2–6 independent experiments. Data points in (J), (K), and (L) represent individual cells. Wilcoxon matched-pairs rank test (F), Kruskal-Wallis test (J, K, L), and multiple unpaired t tests (O) with significance level p < 0.05 were performed for statistical analysis. Scale bars, 50 μm (A) and 10 μm (C, D, E, G, H, and I). See also Figures S2, S4, and S5.