Extended Data Fig. 5: Sensitivity of ZHP-3–ZHP-4 droplets to the high salt concentrations.
From: Crossover patterning through condensation and coarsening of pro-crossover factors
a, Coomassie blue-stained gel exhibiting the purified ZHP-3 and ZHP-4 complex from HEK293T cells. GFP-StrepII-ZHP-3 and His-mCherry-ZHP-4 were transiently coexpressed in HEK293T cells and then purified initially using Ni-NTA agarose beads, followed by a secondary purification step with the Strep-Tactin beads. Approximately 100 ng each of ZHP-3 and ZHP-4 were separated in the lane on the right. b, Representative fluorescence, and Differential Interference Contrast (DIC) images showing condensates formed by 50 nM ZHP-3 (green) and 50 nM ZHP-4 (magenta) complex at various salt and 1, 6-hexanediol concentrations in vitro. Scale bar, 1 μm. c-e, Statistical analyses of ZHP-3–ZHP-4 droplets as described in panel b. c, Droplet number per imaging field. n = 20, 10, 11, 13, 12, 24, 23, 12 imaging fields, for 100 mM, 150 mM, 200 mM, 250 mM, 300 mM NaCl and 5%, 10%, 15% hexanediol, respectively, pooled from two independent replicates. Each imaging field = 2023 μm2. d, Total droplet area per imaging field. n = 20, 10, 11, 13, 12, 24, 23, 12 imaging fields for 100 mM, 150 mM, 200 mM, 250 mM, 300 mM NaCl and 5%, 10%, 15% hexanediol, respectively, pooled from two independent replicates. e, Individual droplet size. Only droplets with a size > 0.1 μm2 were scored due to the resolution limit. n = 2046, 893, 401, 126, 18, 3011, 2772, 1047 droplets for 100 mM, 150 mM, 200 mM, 250 mM, 300 mM NaCl and 5%, 10%, 15% hexanediol, respectively, pooled from two independent replicates. Data are presented as mean ± s.d. for c-e. Ordinary one-way ANOVA, Tukey’s multiple comparisons test with a single pooled variance and each P value adjusted for c and d, and two-tailed Kolmogorov-Smirnov test for e. Source numerical data and unprocessed gel are available in source data.
