Fig. 2

Differentiation and characterization of human cortical neurons and astrocytes derived from FTD-GRN patient and control hiPSCs. a Brightfield images of the cells at different stages of cortical neuronal differentiation. SB = SB 431542, LDN = LDN 193189, RI = Y-27632, BP = BrainPhys™, N2 = N-2 supplement, B27 = B-27 supplement. b Representative immunofluorescence images of DIV 50 WT, S116X^+/−, R418X^+/−, and R493X^−/− KI hiPSC-derived cortical neurons stained for MAP2, GFAP, TUJ1, FOXG1 (forebrain), and TBR1 (cortical layer VI). Cell nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. c-f Cells positive for MAP2, GFAP, FOXG1, and TBR1 (neuronal, astrocytes, forebrain, and cortical layer VI markers, respectively) as a percentage of DAPI⁺ cells. On average, ~ 900 cells were analyzed per replicate, in (c-f) n = 3 independent cultures, 3 images per biological replicate; values are shown as mean ± SEM. g Representative immunofluorescence images of DIV 60+ WT and R493X^−/− KI hiPSC-derived astrocytes stained for GFAP. Cell nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. h Cells positive for GFAP as a percentage of DAPI⁺ cells. On average, ~ 400 cells were analyzed per replicate, n = 3 independent cultures, 3 images per biological replicate; values are shown as mean ± SEM