Figure 4. Zyxin interacts with Siah2 and Lats2.

(a) HeLa cells were transiently transfected with Myc-Lats2 and Zyxin-V5 and subjected to immunostaining with anti-Myc and anti-V5 antibodies. Images were collected by confocal microscopy. R is the colocalization correlation coefficient. Scale bars, 20 μm. (b) Endogenous interaction between Lats2 and Zyxin in MDA-MB-231 cells was analysed by immunoprecipitation. (c) Coimmunoprecipitation of exogenously expressed Myc-Lats2 with Zyxin-V5. (d) Dissection of the critical regions of Zyxin binding to Lats2 in HEK293T cells. (e) Coimmunoprecipitation of exogenously expressed Flag-Siah2^RM with Zyxin-V5. (f) Dissection of the critical regions of Zyxin binding to Siah2 through GST pull-down. Asterisk indicates non-specific binding. (g) Dissection of the critical regions of Siah2 binding to Zyxin in HEK293T cells. (h) Schematic drawing of critical regions of Siah2 and Zyxin and their responses to interaction with Lats2. ‘+' indicates as binding, ‘−' as no binding, and ‘/' as not examined. L, lim domain; RF, ring finger domain; SBD, substrate binding domain.