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. 2021 Sep 23;16(10):2432–2441. doi: 10.1016/j.stemcr.2021.08.014

Figure 1.

Figure 1

Construction and characterization of inducible cell lines

(A) Schematic of PiggyBac transposon-based inducible overexpression vector.

(B) Schematic of cell line generation and validation process.

(C) Inducible NEUROD1 (iN) cell line validation at 3 weeks post induction via (1) qRT-PCR analysis of signature neuronal markers MAP2, TUBB3, VGLUT2, and VGAT; data represent the mean ± SD (n = 4 independent experiments); (2) immunofluorescence micrograph of MAP2+ cells (scale bars, 50 μm); and (3) representative spike plots from MEA measurements of spontaneously firing iN cells.

(D) Inducible ASCL1+DLX2 (iAD) cell line validation at 3 weeks post induction via (1) qRT-PCR analysis of signature neuronal markers MAP2, TUBB3, VGLUT2, and VGAT; data represent the mean ± SD (n = 4 independent experiments); (2) immunofluorescence micrograph of MAP2+ cells (scale bars, 50 μm); and (3) representative spike plots from MEA measurements of spontaneously firing iAD cells.

(E) Inducible MYOD1+BAF60C (iMB) cell line validation at 2 weeks post induction via (1) qRT-PCR analysis of signature skeletal muscle markers MYH8, TNNC1, and RYR; data represent the mean ± SD (n = 3 independent experiments); and (2) immunofluorescence micrograph of MYH+-, MYOG+-, and SAA+-labeled cells (scale bars, 50 μm). (C–E) ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, and ∗∗∗∗p ≤ 0.0001; ns, not significant.