Protocol summary for rupture event localisation and classification.

Each force spectroscopy curve (1) was analysed separately. After contact point fitting, the first derivative of the force was calculated (2). A threshold was set to locate peaks in the derivative (3) correspondent to rupture events (4). A data interval of 50 nm prior to the rupture event was fitted with a line (5); the slope was evaluated to classify the rupture event as cytoskeleton anchored or membrane tether.