This view is consistent with the timing observed experimentally in which one observes active Cdc42 polarization, which is followed by receptor and G-protein polarization (Yi et al., data not shown).
In Figure 4B, we observed that for h = 8 and Lslope = 0.01 µm−1, there was no tracking (but excellent polarization) whereas at Lslope = 0.1 µm−1, the polarization was reduced but the tracking was better (data not shown).
However, to our surprise, INZ or its close analogue INZ1 that induced p53 did not apparently affect the interaction between either MDMX and p53, or MDM2 and MDMX, or MDM2 and p53 in our in vitro fluorescence polarization and cell-based co-immunoprecipitation assays (data not shown).
Moreover, no flotillin polarization was observed during wound healing in these cells (data not shown).
Comparisons of unstained cells to cells stained with only LysoSensor Green or MitoTracker Orange showed no significant changes at the 95% level, except for SiHa cells interrogated with parallel polarization in which the stained cells had decreased scattering (data not shown).
As shown with actin and Gp135 staining, and also with proper localization of ZO-1, a tight junction marker (data not shown), these cells were indeed polarized but failed to exhibit any polarization of flotillins.
Notably, flagellin also promoted Th2 polarization in naïve T cells derived from three nonallergic individuals (data not shown), indicating that the Th2-promoting capacity of flagellin is not restricted to the atopic status.
Dvl2 polarization was not detected at E12.5 in the control digit chondrocytes (data not shown) but emerged at E14.5 appearing as a distally enriched pattern and the opposite of Vangl2 (Fig. 7C and supplementary material Fig. S6C, left panel).
Consistent with these reports, we observed that Vangl2 polarization is first detected in early neurulae at the posterior neural plate (data not shown).
