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Figure 5
In vitro actin co-sedimentation assay (F-actin binding) and negative staining electron microscopy. SDS–PAGE analysis of in vitro actin co-sedimentation assays with EFhd1 and EFhd2. Protein samples (12 µM) were added to polymerized (a) β-actin (8 µM) or (b) α-actin (8 µM) in the presence of 1 mM EGTA, 1 mM CaCl2 or 20 µM ZnCl2. (c) Co-sedimentation ratios from each experiment. Filled and open black squares show the co-sedimentation ratios for EFhd1 and EFhd2 with β-actin, respectively, while filled and open red spheres show the co-sedimentation ratios for EFhd1 and EFhd2 with α-actin, respectively. Symbols and error bars represent the mean and 95% confidence interval for the mean, which were calculated from five independent experiments. Negatively stained electron micrographs: (d) F-actin assembled from β-actin in the presence of EFhd1 and 1 mM EGTA, 20 µM CaCl2 or ZnCl2; (e) F-actin assembled from α-actin in the presence of EFhd2 and 1 mM EGTA, 20 µM CaCl2 or ZnCl2.

IUCrJ
Volume 10| Part 2| March 2023| Pages 233-245
ISSN: 2052-2525
https://doi.org/10.1107/S2052252523001501