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[Preprint]. 2025 Oct 16:2025.10.08.681260. [Version 2] doi: 10.1101/2025.10.08.681260

Figure 3. PINK1-SPARK visualizes PINK1 activity following multiple types of PINK1 activation.

Figure 3.

A. Representative image of HeLa cells expressing PINK1-SPARK when treated with 50 μM kinetin riboside (KR).

B. Number of puncta per cell in WT or PINK1 KO HeLa expressing PINK1-SPARK treated with DMSO (WT: n = 118 cells from 3 experiments; PINK1 KO: n = 236 cells from 3 experiments) or 50 μM KR (WT: n = 129 cells from 3 experiments; PINK1 KO: n = 169 cells from 3 experiments; ****p < 0.0001; ns = 0.2897; unpaired t-test, two-tailed).

C. Representative image of HeLa cells expressing PINK1-SPARK when treated with MTK458 (3.1 μM) and CCCP (0.5 μM).

D. Number of puncta per cell in WT or PINK1 KO HeLa expressing PINK1-SPARK when treated with DMSO (circles, WT: n = 69 cells from 3 experiments; PINK1 KO: n= 236 cells from 3 experiments) or 3.1 μM MTK + 10 μM CCCP (diamonds, WT: n = 81 cells from 2 experiments; PINK1 KO: n = 201 cells from 3 experiments; ****p < 0.0001; unpaired t-test, two-tailed).

E. Comparison of puncta size between treatment conditions: 10 μM CCCP (n = 481 puncta from 3 experiments), 3.1 μM MTK + 0.5 μM CCCP (n = 1451 puncta from 3 experiments), or 50 μM KR (n = 4727 puncta from 3 experiments; ***p = 0.0002; unpaired t-test, two-tailed).

For all images, scale bars represent 10 μm. Bar graphs display mean ± standard deviation.