Fig. 2: Colocalization between Gag-mEos4b and AF647-NT-Lys in PALM/dSTORM imaging.

a Confocal images of Gag/Gag-mTagBFP2 derivatives-expressing HeLa cells labeled with EGFP-NT-Lys and mCherry-D4. Bar, 10 µm. b, g PALM/dSTORM images of Gag/Gag-mEos4b derivatives-expressing cells labeled with AF647-NT-Lys (b) and AF647-D4 (g). Bar, 5 µm c, d, h, and i Regions enclosed by yellow rectangles in (b) and (g) are enlarged. c Images in a cell expressing Gag WT/Gag WT-mEos4b and labeled with AF647-NT-Lys. The yellow dotted lines indicate a similar pattern of labeling with both fluorophores. d Images in a cell expressing Gag-WM/Gag-WM-mEos4b and labeled with AF647-NT-Lys. The yellow arrows indicate Gag-WM-mEos4b signals that were not associated with AF647-NT-Lys signals (see text). h Images in a cell expressing Gag WT/Gag WT-mEos4b and labeled with AF647-D4. i Images in a cell expressing Gag-WM/Gag-WM-mEos4b and labeled with AF647-D4. The yellow arrows indicate Gag-WM-mEos4b signals that looked independent of AF647-D4 signals. Bar, 1 µm. e, f Mander’s overlap coefficient (MOC) values for colocalization of Gag WT-mEos4b (WT) and Gag-WM-mEos4b (WM) to AF647-NT-Lys (e) and of AF647-NT-Lys to WT and WM (f). j, k MOCs of WT and WM to AF647-D4 (j) and of AF647-D4 to WT and WM (k). In the graphs, dots and bars in three different blue and black bars with error bars indicate means in each cell, means in each experiment, and means ± SEM in three independent experiments (n = 18 for WT and 16 for WM over three experiments). * and ** in the graphs indicate significant differences at significance levels of 0.05 and 0.01 in the paired two-tailed t-test. Source data and p-values for (e, f, j, and k) were provided as a Source Data file.