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. Author manuscript; available in PMC: 2017 May 1.
Published in final edited form as: Biotechnol J. 2016 Feb 16;11(5):662–675. doi: 10.1002/biot.201500374

Figure 4.

Figure 4

YAP knockdown decreased YAP levels and transcriptional activity. (A) Schematic of construct for dox-inducible shRNA knockdown of YAP expression. PH1-TetO is the Tet-responsive human H1 promoter. H9 hESC YAP knockdown and scrambled sequence shRNA (ishscram) cell clones were plated at 0.2 ×105 cell/cm2 and treated with 2 µM doxycycline (dox) for 3 days and (B) harvested for qPCR analysis of YAP expression (• indicates p<0.05 compared to ishscram control) or (C) harvested for western blot analysis of YAP protein levels. (D) After 3 days of dox treatment, cells were transfected with the YAP responsive 8xGTIIC firefly luciferase plasmid. Following an additional 24 hours of dox treatment, cells were harvested for a luciferase assay. (• indicates p<0.05 compared to no dox condition) (E) Cells were treated with dox for 3 days and harvested for PCR analysis of YAP target genes, YAP, TAZ, POU5F1 (Oct4), NANOG and SOX2.