Visualizing INVs by light microscopy. (A) Single ROIs from live-cell imaging experiments with cells expressing GFP or GFP-TPD54 (overexpressed [OE]), or with knock-in GFP-TPD54 cells (endo). Images are pseudocolored to highlight subresolution structures. Scale bar, 1 µm. (B) Scatter dot plot to show the mean variance per pixel over time. Dots, individual cells; black bars, mean ± SD. The mean ± SD for GFP is indicated as a thin black line and gray zone. n_cell = 16–20. (C) TIRF image showing the ventral surface of a typical GFP-TPD54 knock-in HeLa cell imaged by STORM. Scale bar, 10 µm. (D) Expanded view of the boxed region in C. Scale bar, 500 nm. (E) STORM image of the corresponding region shown in D. Localizations are pseudocolored as indicated; max value in image was 79. Scale bar, 500 nm. (F) Histogram of FWHM values of all spots in the entire localizations image for the cell shown in C. (G) Summary of median FWHM values. Bars, mean ± SD. n_cell = 5, n_exp = 3.