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. 2020 Dec 15;31(26):2932–2947. doi: 10.1091/mbc.E19-10-0591

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© 2020 Shanmugam 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 5: — UNC-104 clustering is increased in ptp-3 mutants and decreased in syd-2 mutants. (A) UNC-104::mRFP accumulations and distribution patterns in straightened segments of sublateral neurons taken from different genetic backgrounds stacked on each other with: control = unc-104(e1265) allelic mutants rescued with UNC-104::mRFP; ptp-3(ok244) = ok244 allelic mutants expressing UNC-104::mRFP; ptp-3(mu256) = mu256 allelic mutants expressing UNC-104::mRFP; ptp-3(mu256) rescue = mu256 allelic mutants expressing both PTP-3B::CFP and UNC-104::mRFP; syd-2(ok217) = ok217 allelic mutants expressing UNC-104::mRFP; syd-2(ok217);ptp-3(mu256) = allelic double mutants expressing UNC-104::mRFP; syd-2(ok217) rescue = ok217 allelic mutants expressing both GFP::SYD-2 and UNC-104::mRFP; syd-2(ok217)[SYD-2 Y741F] = ok217 allelic mutant expressing both nonphosphorylatable GFP::SYD-2 Y741F and UNC-104::mRFP; syd-2(ok217)[SYD-2 Y741E] = ok217 allelic mutant expressing both phosphomimicking GFP::SYD-2 Y741E and UNC-104::mRFP. (B) UNC-104 cluster sizes in sublateral neurons. (C) UNC-104 densities along sublateral neurons. (D) Representative immunoblots from co-IP of UNC-104 from phosphodeficient (Y741F) and phosphomimicking (Y741E) SYD-2. (E) Quantification of coprecipitated UNC-104 either with phosphodeficient or phosphomimicking SYD-2. Scale bar: 10 µm. Box plot with median and error bars: ± max. and min. range. One-way ANOVA with Fisher’s LSD multiple comparison test. *p < 0.05, **p < 0.005, ***p < 0.001, and ****p < 0.0001. F (DFn, DFd) = 7.073 (8, 238) (C); 6.543 (2,6) (E).