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. 2011 May 16;193(4):633–642. doi: 10.1083/jcb.201009069

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© 2011 Habbig et al.

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Figure 1. — NPHP4 interacts with Lats1 and activates TAZ/YAP/TEAD transcription. (a) HEK293T cells were transiently transfected with the FLAG-tagged NPHPs NPHP1 (F.NPHP1) or NPHP4. After immunoprecipitation (IP) with FLAG (M2) beads, Western blot analysis revealed that endogenous Lats1 coprecipitated with NPHP4 but not with NPHP1. (b) Lysates from HEK293T cells transfected with either control vector (Input 1) or FLAG-tagged NPHP4 (Input 2 + 3) were subjected to immunoprecipitation using an anti-Lats1 (IP 1 and 2) or a control (anti-V5; IP 3) antibody. Western blot (WB) analysis showed that NPHP4 coprecipitated specifically with endogenous Lats1. (c) NPHP4 enhances the activity of the transcriptional coactivators TAZ and YAP. Plasmids encoding the indicated proteins or empty pcDNA6 vector were cotransfected in HEK293T cells together with the TEAD reporter plasmids as indicated in Materials and methods. Coexpression of NPHP4 increased the TAZ/YAP-dependent signaling to 156 and 138%, respectively (n = 3 for YAP and n = 5 for TAZ; *, P < 0.05). Expression of the indicated proteins was confirmed by Western blot analysis. (d) HEK293T cells were transfected with the indicated constructs. The chromatin immunoprecipitation assay revealed that NPHP4 enhances the binding of TAZ to the promoter region of CTGF, a known target gene of TAZ. Error bars represent SEM.