Figure 1.

Sox9, Nfia, and Nfib convert human embryonic fibroblasts to induced astrocytes

(A) Schematic of doxycycline-inducible lentiviral constructs.

(B) Experimental overview of direct conversion to induced astrocytes (iAs).

(C) Representative immunofluorescence images of GFAP and S100B in rtTA- and SAB-infected HEFs at 3 weeks.

(D) Quantification of S100B, GFAP, and S100B/GFAP immunoreactive cells at 3 weeks related to total number of nuclei (purity) and plated cells (yield).

(E) Representative immunofluorescence images of SAB-HEF-iAs at 3 weeks converted in serum-containing medium (+Serum), serum-free medium (−Serum), serum-free medium with small molecules (−Serum + SM), or low-serum-containing medium with SM and FGF2 followed by serum-free medium with SM (FGF2 − Serum + SM).

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

(G) Immunofluorescence image of SAB-HEF-iAs at 5 weeks converted in FGF2 − Serum + SM medium.

Puro, puromycin resistance gene; Blast, blasticidin resistance gene; Hygro, hygromycin resistance gene; HEFs, human embryonic fibroblasts. Individual color levels are adjusted for visibility in (C), (E), and (G). Data are presented as mean ± SEM of n = 3–4 independent experiments. Two-tailed paired t tests (D) and Kruskal-Wallis test (F) were performed for statistical analysis. ^∗p < 0.05, ^∗∗p < 0.01. Scale bars, 100 μm (C), 25 μm (E), 50 μm (G), and 20 μm (G, inset).