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. 2019 Dec 18;13:535. doi: 10.3389/fncel.2019.00535

Figure 5.

Figure 5

Analysis of PDE activity from individual cells reveals distinct PDE activity profiles. (A) Schematic of PDE activity profiles from individual cells revealed three distinct cAMP decay profiles, rapid, intermediate, and delayed. (B) PBM index score of cAMP degradation profile clusters in HEK293T and SH-SY5Y cells. Data from control cells or cells expressing WT or a mutant variant of α-synuclein were merged for analysis. A maximum at three clusters indicates cAMP degradation profiles can be best segmented into three distinct profiles for both HEK293T and SH-SY5Y cells. (C) Individual cAMP degradation profiles from control HEK293T cells and HEK293T cells expressing WT or a mutant variant of α-synuclein. Each black line represents a single cell; the red line represents the mean response of each profile. N = 3 wells per sample, control = 4,047 cells, WT = 3,882 cells, A53T = 3,385 cells, E46K = 3,431 cells, A30P = 3,700 cells, H50Q = 3,455 cells. (D) Bar plot of the fraction of cells in each profile for control and α-synuclein variants from the profiles determined in (C). N = 3 wells for each condition. Individual data points are plotted along with the mean bar and error bars representing the standard deviation. Panels (E,F) same as in (C,D) but for SH-SY5Y cells. N = 3 wells per sample, control = 2,697 cells, WT = 2,427 cells, A53T = 2,775, E46K = 1,462 cells, A30P = 1,902 cells, H50Q = 2,209 cells.