Figure 4: YAP/TAZ inhibit ciliogenesis and upregulate the expression of AURKA and PLK1.

(a) Immunoblotting to confirm siRNA-mediated knockdown of YAP and TAZ. RPE1 cells were transfected with the indicated siRNAs for 60 h. Transfection of YAP si2 depleted both YAP and TAZ. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. (b) Quantification of ciliogenesis in cells transfected with YAP, TAZ and YAP/TAZ mix siRNAs for 60 h in the presence of 10% FBS. (c) Graph of ciliated cell frequency in cells co-depleted with YAP and TAZ in the absence of serum. LPA (30 μM) was added to the culture media 24 h before fixation. (d) Quantification of ciliogenesis in cells infected with retroviruses carrying an empty vector, YAP-5SA (a hyperactive YAP mutant) or YAP-5SA-S94A (a hyperactive YAP mutant without TEAD-binding capacity), and then cultured in serum-free media for 48 h. (e) Quantitative RT–PCR (qRT–PCR) analyses of the expression of AURKA and PLK1 in cells transfected with YAP/TAZ mix siRNAs for 60 h. GAPDH was used as a normalization control. (f) qRT–PCR analyses of the expression of AURKA and PLK1 in cells infected with retroviruses carrying an empty vector, YAP-5SA or YAP-5SA-S94A, and then cultured in the presence of serum for 48 h. (g) Graph showing the ciliated cell percentage in cells transfected with the AURKA and PLK1 siRNAs for 60 h. DMSO or CytoD was added to the culture media 6 h before fixation. All error bars are s.d. (n=3 independent experiments). The P-values were determined by unpaired Student’s t-test; *P<0.05, **P<0.01.