Figure 3. Mechanical strain induces a Src-dependent increase in Y654 phosphorylated β-catenin and β-catenin transcriptional activity.
Distribution of TOPdGFP at 8 hr (A), β-catenin at 8 hr (A, insets), EdU at 24 hr (C), and pY654 β-catenin at 8 hr (E) in MDCK monolayers after No Strain or High Strain (15%) applied by the ISA, treated with either DMSO or the Src inhibitor SU6656 (10 μM). Scale bars: 25 μm. Quantification of percent cells TOPdGFP- (B) or EdU- (D) positive and quantification of average pY654 β-catenin intensity per pixel (F); note that the small surface area (0.81 cm2) of the ISA does not provide sufficient cell numbers for biochemical characterization. Quantifications were from at least 3 independent experiments and for the TOPdGFP and EdU quantifications included analysis of 677–1168 cells per experiment. Quantifications were mean +/- SEM; unpaired t-test (B,D) or Kolmogorov-Smirnoff (F) test p values<0.05 (*), <0.01 (**), and <0.001 (***).
DOI: http://dx.doi.org/10.7554/eLife.19799.011
Figure 3—figure supplement 1. Distribution of β-catenin in monolayers treated with either DMSO or the Src Inhibitor SU6656 8 hr after the application of No Strain or High Strain (15%) using the ISA.
Figure 3—figure supplement 2. The Src Inhibitor PP2 blocks strain-induced increases in Y654 phosphorylated β-catenin and β-catenin transcriptional activity.
Figure 3—figure supplement 3. Distribution of β-catenin in monolayers treated with either DMSO or the Src Inhibitor PP2 8 hr after the application of No Strain or High Strain (15%) using the ISA.
Figure 3—figure supplement 4. EGFR inhibition reduces an increase in pY654 β-catenin following mechanical strain, but does not affect β-catenin transcriptional activity or cell cycle re-entry.
Figure 3—figure supplement 5. Distribution of β-catenin in monolayers treated with either DMSO or the EGFR Inhibitor PD153035 8 hr after the application of No Strain or High Strain (15%) using the ISA.
