Fig. 5

The IQGAP1-Hax1 interaction regulates cell migration. A, In MCF7 cells, suppression of Hax1, IQGAP1, or Hax1/IQGAP1 exhibits the same defect in directional migration. The migration of confluent monolayers of mock, Hax1 -depleted, IQGAP1-depleted, or Hax1/IQGAP1-depleted MCF7 cells were scratched, followed by visualization with phase-contrast microscopy at the indicated time points. B, Characterization of the efficiency and specificity of various siRNA oligonuclotides in MCF7 cells. The protein levels of Hax1, IQGAP1, or Hax1/IQGAP1 were significantly decreased in MCF7 cells transfected with corresponding siRNAs. C, The kinetics of in vitro wound healing in A are quantified. Note that suppression of Hax1, IQGAP1 or Hax1/IQGAP1 in MCF7 cells all lead to significant delay of in vitro wound healing (n = 3, p < 0.0001, Student’s t test). D, In MCF7 cells, directional migration of Hax1 knockdown cells and Hax1 –depleted cells rescued with different truncation mutants of Hax1 were evaluated by in vitro scratch wound assays. The migration of confluent monolayers of mock, Hax1 -depleted, or MCF7 cells co-transfected with Hax1 siRNA and two HA-Hax1 constructs (HA-Hax1-NT and HA-Hax1-CT) were scratched, followed by visualization with phase-contrast microscopy at the indicated time points. E, The kinetics of in vitro wound healing in D are quantified. Note that re-expression of Hax1-CT but not Hax1-NT mutant in Hax1-depleted MCF7 cells successfully rescues the defect of in vitro wound healing (n = 3, p < 0.01, Student’s t test)