fig3

Figure 3. EGF-driven cell migration is inhibited in paxillinS178A cells because of impaired focal adhesion turnover. MTLn3 cells were either untreated or treated with EGF (10 nmol/L). A: migration was observed for 10 h by epi-fluorescence microscopy PaxillinS178A reduced cell speed average (about 100 cells per condition were imaged in one biological replicate). This graph shows the data for one representative biological replicate.*P < 0.05, **P < 0.01, ***P < 0.001 (a) and directional cell movement (b). See also Supplementary movie M2; B: matrix adhesions dynamics in MTLn3 cells was visualized with TIRF microscopy. See alsoSuplementary movie M3. Overlay of different timeframes were generated in red, green and blue. Focal adhesions in white, as observed for PaxillinSer178, represent unchanged (less dynamic) focal adhesions. Scale bar is 20 μm; C: protein dynamics was measured with the spot bleaching technique and showed similar dynamics for both GFP-paxillinS178A (b) and GFP-paxillin-wt (a). Approximately 20 focal adhesions (each in distinct cells) were averaged to generate one FRAP curve for a single experiment.The mean relative fluorescence of both GFP-paxillin-wt and GFP-paxillinS178A both in SFM and upon EGF stimulation shows no significant difference after 30 s of recovery