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. Author manuscript; available in PMC: 2018 Dec 26.
Published in final edited form as: Lab Invest. 2018 Jun 26;98(9):1126–1132. doi: 10.1038/s41374-018-0068-x

Figure 3. Signaling in NKX2-1-overexpressing cells.

Figure 3

(a) GFP immunofluorescence to determine the efficiency of NKX2-1 overexpression in the presence or absence of doxycycline (Dox, 1 µg/ml) in SPTL cells. (b) Western blotting for thyroid and epithelial differentiation markers in SPTL cells cultured for 2 days under 0.1% FBS without (−) and with (+) Dox (1 µg/ml). Thy: mouse thyroid follicular cells. β-actin was used as a loading control. (c) Cell morphology of SPTL cells cultured under DOX. Images were taken after 3 days culture under 2 % FBS in the absence or presence of Dox (3 µg/ml). (d) Western blotting for DOX dose-dependent expression of NKX2-1, ERK, pERK, AKT, and pAKT in SPTL cells. Cells were cultured under 0.1% FBS and treated with DOX for 2 days. 10% FBS was used as positive control for pAKT. α-tubulin was used as a loading control. (e) Immunofluorescence for NKX2-1 and pAKT in a zone of cells located near the tracheal cartilage and muscle in Nkx2-1(fl/fl);TPO-cre mouse. Arrows: cells expressing both pAKT and NKX2-1.