Fig. 6: ALG2 and CALC alter the nanomechanical properties of ANXA11-GUV assemblies and their ability to tether RNP granules.

a A Schematic of our microfluidic device used to extract the relative elastic modulus of GUVs. The bright-field images on the right illustrate GUV deformation within the device with a channel size: opening = 15 µm, tapered end = 2 µm. b A plot of the GUV deformation (strain) under variable pressure applied across the V-shaped channel (stress). Simple linear regression, R² (Forth/Back) – 0.991/0.980. ANCOVA-slopes (p = 0.26) and intercepts (p = 0.13) are not significantly different. c Quantification of the relative elastic modulus of GUVs at 100 µM Ca²⁺ with 0.5 µM ANXA11 FL, coincubated with either 2 µM ALG2 or 20 µM CALC. Mean ± SD. One-way ANOVA with Tukey’s multiple comparison, **p = 0.0023, ****p < 0.0001, n = 3 GUVs. d The experimental pipeline for FAPS-based RNP granule isolation from a stable G3BP1-mEmerald U2OS line. Created in BioRender. Nixon-Abell, J. (2025) https://BioRender.com/j66j626. e Representative fluorescence images of ATTO594 GUVs incubated with 100 µM Ca²⁺ and 0.5 µM ANXA11 co-incubated with either 2 µM ALG2 or 20 µM CALC. To each condition, purified RNP granules (labelled with mEmerald-G3BP1) were added to a final concentration of 0.2 mg/ml. Scale bar - 5 µm. f Quantification of the fluorescence intensity of RNP granules (mEm-G3BP1) recruited to ANXA11-GUV assemblies as displayed in (e). Mean ± SD. One-way ANOVA with Tukey’s multiple comparison, *p = 0.0111 (A11 vs A11 + ALG2) ;0.0125 (A11 vs A11 + CALC), ****p < 0.0001, n = 7 repeats (21–78 GUV).