Fig. 4
From: Regulation of Yki/Yap subcellular localization and Hpo signaling by a nuclear kinase PRP4K
PRP4K plays a conserved role in mammalian Hpo signaling pathway. a, b Wild type or Lats1/2 −/− HEK293A cells were transfected with the indicated constructs. Cell extracts were separated on phos-tag conjugated or regular SDS-PAGE and immunoblotted with the indicated antibodies. c HEK293A transfected with the indicated constructs were immunostained with YAP and GFP antibodies and DAPI. d HEK293A cells were transfected with control or PRP4K siRNA in the absence or presence of PRP4K/PRP4KKR-GFP and starved for 6 h prior to lysis. Cell extracts were separated on phos-tag conjugated or regular SDS-PAGE and immunoblotted with the indicated antibodies. e HEK293A cells were transfected with control or PRP4K siRNA and starved for 6 h before fixation and immunostaining with Yap and tubulin antibodies. f Wild type and Lats1/2−/− HEK293 cells were transfected with control or PRP4K siRNA and starved for 6 h prior to lysis. Cell extracts were separated on phos-tag conjugated or regular SDS-PAGE and immunoblotted with the indicated antibodies. g Schematic drawing of Yap with Lats1/2 phosphorylation sites aligned underneath. TBD: TEAD binding domain. WW: WW domain. AD: activation domain. h–k Cell extracts from HEK293 cells transfected with the indicated constructs were separated on phos-tag conjugated or regular SDS-PAGE and immunoblotted with the indicated antibodies. l 3XSd2-luc reporter assay in HEK293 cells transfected with the indicated constructs. Data are means ± s.d. from three independent experiments. m, n HEK293 cells were transfected with the indicated constructs. Cell lysates were immunoprecipitated with a GFP antibody, followed by western blot analysis Flag and GFP antibodies, or directly subjected to western blot analysis with Flag and HA antibodies. Scale bars, 10 μm (c, e)
