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. 2023 Jan 16;9:4. doi: 10.1038/s41531-023-00444-w

Fig. 4. Activity-induced pS129 is enriched in membrane fractions.

Fig. 4

a In vitro kinase assay was carried out by incubating 2 µM recombinant αS alone, αS + 2 µM Plk2, and αS + Plk2 + 0.2 mM liposomes. N = 3 independent experiments on different days, n = 12 biological replicates total; ****p < 0.0001. Mean +/− SD. RM one-way ANOVA with post hoc Sidak’s multiple comparisons test. b DIV18-20 rat cortical neurons treated with 20 µM PTX for the indicated time points and subjected to sequential extraction to generate cytosol (C) vs. membrane (M) protein lysates. WB for total αS and pS129. Quantification of total αS and pS129 in membrane fractions. N = 4 independent experiments on different days, n = 16 biological replicates total. ****p < 0.0001. Mean +/− SD. Two-way ANOVA with post hoc Tukey’s multiple comparisons test. c Samples as in b. Quantification of total αS vs pS129 ratios as indicated in cytosolic and membrane fractions, respectively. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Mean +/− SD. Brown-Forsythe and Welch ANOVA with Dunnett’s T3 post hoc test for multiple comparisons. d DIV18-21 rat cortical neurons treated with DMSO or 20 µM PTX followed by isolation of synaptosomes. WBs for total αS and pS129. N = 6 independent experiments on different days. *p < 0.04. Mean +/− SD. Ratio paired t-test; two-tailed.