Cumulative density histograms of track displacements before and after capsaicin treatment. HEK293T cells expressing TRPV1exCellHalo are labeled with Alexa488 HaloTag ligand for single molecule tracking movies (see Materials and methods). 2.5-s movies (50 frames) are acquired, 15 s apart, before and after 5 μM capsaicin treatment to assess changes in TRPV1exCellHalo mobility. Cumulative density histograms (blue: pretreatment data; red: post-treatment data; purple: overlapping region in histograms) of track displacements are shown for three separate treatment conditions: (top row) capsaicin treatment with TRPV1exCellHalo expressing cells that are replated the same day onto poly-D-lysine coated coverslips; (middle row) mock treatment of TRPV1exCellHalo expressing cells replated onto coverslips the same day; (bottom row) mock treatment of TRPV1exCellHalo expressing cells replated onto coverslips 1 day earlier. A guideline at 0.75 is meant to help observe the upper-quartile values of the displacement distributions. Panels on the far left illustrate ROI-bounded tracks from cells that are paired with data in the blue histogram in the adjacent panel. Summary plots for these data are shown in Fig. 5 D.