Generation of local thickness. Demonstration of how EB1 thickness is calculated from EB1 microtubule tracks (GJA1-WT and GJA1-43k images shown are from Fig. 6 A). Representative images from HeLa cells transfected with either GJA1-WT or GJA1-43k to demonstrate how thickness is calculated from EB1 microtubule tracks. The Trackmate plugin was used to track EB1 particle movements (gray). Next, the Local Thickness plugin generates a distance map (center panel) and then the distance ridge is calculated by removing redundant points from the thickness map. The function Distance Ridge to Local Thickness was then performed to create a distance map of the thickest sections of the image (right panel). Intensity was measured from the right panel images to quantify EB1 microtubule track thickness.