Fig. 5.

GATA3 overexpression in NHB cells. (A) GATA3 overexpressing and control (empty vector) NHB cell cultures following exposure to the PPARγ-activating TZ/PD protocol for 72 h. Western blotting of whole protein lysates was performed to assess protein expression of GATA3, FOXA1 and PPARy1. NHU cells (non-transduced) and treated with the TZ/PD protocol for 72 h are shown for comparison. (B) GATA3 overexpressing and control (empty vector) NHB cells at 72 h post TZ/PD protocol. GATA3, FOXA1 and PPARy protein expression assessed by indirect immunofluorescence microscopy. NHU cells (non-transduced; 72 h TZ/PD protocol) were included as positive controls for comparison. Scale bar ≡ 50 µm. (C) GATA3 overexpressing and control (empty vector) NHB cells were induced to form cell sheets using 5% ABS and 2 mM calcium for up to 7 days. Expression of the tight junction-associated proteins, claudin 3, 4, 5 and 7, assessed by western blotting. ACTB was included as a loading control. NHU cells (non-transduced) exposed to the same protocol were used as a positive control for comparison. Experiments were performed on n = 2 independent NHB donor cell lines and representative results shown.