Figure 2.

Nkx2-1 Overexpression at AFE Stage Results in Efficient Thyroid Differentiation

(A) Experimental schematic of the extended culture conditions in (B), (C) (top three panels), and (D).

(B) RT-qPCR for thyroid marker expression. Fold changes calculated relative to undifferentiated cells, error bars represent SEM (n = 5 independent experiments, n = 1 control).

(C) Immunostaining of induced cultures at D14, D22 (gelatin substratum), and D30 (Matrigel embedded). Scale bars represent 100 μm.

(D) Intracellular flow cytometry for populations at D14 (top), separate differentiation single stains at D22 (bottom).

(E) Schematic of the maturation culture conditions for (F) and (G).

(F) T4 ELISA from D50 (Dox D6–D7) cells ± 10 μM NaI from D40 to D50 (n = 3 wells from the same differentiation). Mouse thyroid tissue for reference (n = 2 tissue samples).

(G) D50 immunostaining for +NaI cultures corresponding with (F). Scale bar represents 100 μm.

^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001. See also Figure S2.