Fig. 5: The activities of 2DGels on F-actin. | Communications Biology

Fig. 5: The activities of 2DGels on F-actin.

From: Structural and biochemical evidence for the emergence of a calcium-regulated actin cytoskeleton prior to eukaryogenesis

Fig. 5

a–d SDS PAGE analysis of high speed sedimentation of F-actin (8 μM) treated with Asgard 2DGels. Soluble (s) and pellet (p) fractions are indicated. The position of actin migration is highlighted by black arrows. a MKD1-2DGel (MKD1, red arrow). Actin, actin alone; MKD1, MKD1-2DGel alone; Ca, 1:1 ratio in 1 mM Ca2+; and EGTA, 1:1 ratio in 1 mM EGTA. The entire SDS PAGE gels are shown in Supplementary Fig. 4. b Heim2DGel (Heim, green arrow). c MKD1-2DGel domain 2 (D2, blue arrow). d Loki2DGel (Loki, purple arrow). e The effect of Loki2DGel on actin filaments. Actin was polymerized in 1 mM EGTA or 1 mM CaCl2 for 10-20 mins. The time course follows the behavior of filaments on adding Loki2DGel (32 μM), followed by TIRF (Supplementary Movie 1). Similar time course for MKD1-2DGel (10 μM) in (f) 1 mM EGTA or (g) 1 mM CaCl2. Arrows indicate severing. h Two time points for the assembly of F-actin (0.4 μM) in the presence of Heim2DGel (32 μM), in 1 mM Ca2+ or 1 mM EGTA, imaged by TIRF microscopy (Supplementary Movie 3). i The apparent elongation rate of individual actin filaments from (h).

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