Supplementary Figure 4: STED/sptPALM correlative nanoscale imaging of focal adhesion sites and adhesion molecules in fibroblasts.

a) From left to right: STED image of paxillin, sptPALM trajectories of integrins in fibroblasts and overlay of both, scale bar 2 µm. b) Zoom of focal adhesion sites which are better resolved by STED (right) compared to widefield (left) and confocal (middle), scale bar 1 µm. c) STED/sptPALM overlay illustrates that immobilized molecules (cyan) colocalize with adhesion sites resolved by STED microscopy, while freely moving molecules (green) are more localized outside the adhesion sites. One representative image (a, b and c) out of 4 independent experiments. d) In widefield and confocal microscopy, immobile trajectories are 10% and 4.5 % underestimated in FAs, respectively, compared to STED (One-way ANOVA, p<0.0001, N = 22 FAs, from 4 cells, 2 cultures), box plots represents the average with range.