Redistribution of endogenous NMIIB in COS-7 cells expressing different levels of GFP-NMIIA. (A–C) Cells with low (A), medium (B), and high (C) expression of GFP-NMIIA (red) fixed and stained with phalloidin and NMIIB antibody (cyan). Bar, 20 µm. (D) Two examples of NMIIB intensity histograms obtained from raw 16-bit images of cells with (right) or without (left) GFP-NMIIA expression. Red lines indicating 1st to 99th percentiles mark the histogram width. (E) Correlation between GFP-NMIIA expression levels (y axis) and the width (1st–99th percentile) of the NMIIB immunofluorescence intensity histogram (x axis; n = 50 cells).