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. 2019 Feb 1;30(3):333–345. doi: 10.1091/mbc.E18-06-0402

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© 2019 Wu et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 3: — Brightness analysis of MEG-3::meGFP SD particles. (A) Representative near-TIRF images of MEG-3::meGFP in mex-5/6(RNAi) embryos before and after bleaching. The overall intensity of the embryo is brighter before bleaching because of higher particle density and out-of-focus signal. Scale bar = 5 μm. (B) Relative number of MEG-3::meGFP particles normalized to the average number between 0 and 1 s and plotted over time. After 100 s of photobleaching, the number of detectable MEG-3::meGFP particles dropped by an average of ∼80%. Average of five embryos. Error bars indicate SEM. (C) Brightness analysis of endogenously tagged MEG-3::meGFP particles in mex-5/6(RNAi) embryos before and after bleaching. Frequency of normalized intensity of all detectable MEG-3::meGFP particles pooled from the beginning of movies (0–1 s, the solid line) or from the end of the movies (95–100 s, the dashed line). Intensity is normalized to the peak value of 95–100 s for individual embryos as described in the methods. n = the total number of particle brightness estimates from five embryos. Note that because we did not track individual particles between frames, the brightness of each particle was estimated in each frame in which it was detected.