FIG 8.

AJUBA LIM proteins do not inhibit Hippo activation of LATS at the plasma membrane. (A and B) HEK293T cells were transfected with MST2, LATS2, LIMD1, and either wt MOB1A (A) or mp-MOB1A (membrane-targeted MOB1A) (B). MOB1A, LATS2, and LIMD1 immunofluorescence assays were performed, as indicated. Nuclei were visualized with DAPI stain. Scale bars, 50 μm. (C and D) HEK293T cells were transfected with MST2, LATS2, or WW45, with or without LIMD1 and with either MOB1A or mp-MOB1A, as indicated. (C) The cells were lysed and Western blotted with the indicated antibodies (input control, 10% of the cell extracts used for IP). (D) MST2 was immunoprecipitated, and the bound products were Western blotted with the indicated antibodies. The amount of pT1041.LATS2 detected is shown below each lane in the top panel. The amount of pT1041.LATS2 present in cells not transfected with LIMD1 was arbitrarily set as 1. All pT1041.LATS2 levels were normalized to total LATS2.