Fig.4.

G-/F-actin spin-down assay. A) Analysis of G- and F-actin enriched fractions from LCLs. RIPA lysates were subjected to spin-down fractionation as described in Fig. 1 and in the Materials and Methods. The resulting G-actin enriched supernatant (S) and F-actin enriched pellet (P) fractions were analyzed by immunoblotting for α-tubulin and β-actin. To visualize protein bands, a parallel gel was prepared and stained with Coomassie Blue (CB below). β-actin bands are marked. B) Effect of incubating with ATP and MgCl₂ on electrophoretic patterning of actin. Actin pattering conversion was performed as described in the Materials and Methods. +ATP: incubated ATP and MgCl₂; – ATP: untreated mock sample. Human platelets G-actin standard (AcSt) (Cytoskeleton Inc.) was included for comparison. C) Spin down assay of Lewy body disease (LBD) and frontotemporal lobe dementia (FTLD) LCLs. Details were as described in (A). D) Spin-down assay of pools LCLs lysates. Each pool was prepared by combining three distinct LCLs lysates and subjected to spin-down assay probing for DYRK1A, α-tubulin, and β-actin as described in (A).