Actin disruption leads to loss of GPI-GFP periodicity. (a) Correlative SPT of GPI-GFP of progenitor-derived neuronal cells before and after SWIN A treatment. GPI-GFP was tracked using anti-GFP-NB-conjugated QDs (5,000 frames, 200 Hz). Cells were then treated with 1 µM SWIN A for 30 min. Afterward, the same cell was tracked again. A local “stripe score” was then calculated based on the localizations via a Fourier-filter-based algorithm to detect periodicity at 200 nm. Shown are reconstructions of SPT experiments overlayed with heat maps of the local stripe score. After treatment, cells show a clear reduction in periodicity at 200 nm. Scale bar is 5 µm. (b) Heat map of stripe score of box in (a) before and after SWIN A treatment. (c) Reconstructions of SPT experiments of box in (a) before and after SWIN A treatment. (d) Autocorrelation function (ACF) of periodic area in (c) as indicated with dotted lines before and after SWIN A treatment. Periodicity ∼200 nm is lost after SWIN A treatment. (e) Experimental procedure as described in (a). Violin plot of stripe scores of cells before and after SWIN A treatment (n = 30 per condition from three independent experiments). Lines indicate means. Normality was tested using Shapiro–Wilk test and rejected (DMSO after P = 0.072 ns, SWIN A after P = 0.022*). Significance was tested using Mann–Whitney test. SWIN A treated cells show a significantly reduced stripe score compared to the control cells (DMSO after versus SWIN A after P = 4.2E-10****). Scale bars are 500 nm if not otherwise indicated.