Fig. 5: Delivered VHHGFP4 HBpep-SA “Hubs” bind to and enrich native target GFP protein.
From: Delivery of peptide coacervates to form stable interaction hubs in cells
A, B Representative fluorescence images of target binding to hubs inside U2OS cells stably expressing GFP, comparing mCherry control and VHHGFP4(nanobody)mCherry-loaded coacervates. 30 µl of coacervates formed from 0.5 mg/mL HBpep-SA, co-assembled with 0.1 mg/mL mCherry or VHHGFP4-mCherry were added to the cell media for U2OS cells and imaged at 24 h. Cell boundaries are indicated by dashed gray line. Chosen “hub” for line scan is indicated by white line. Scale bar: 10 µm. C, D Line scans of coacervate hubs showing fluorescence enrichment in (A, B). E Quantification: percentage of hubs in cells that show max enrichment index greater than 2. Sample size n = 902, 865, 1145, 986. F Quantitation of percentage of cells that show at least one hub recruiting target with EI > 2. N = 66, 68, 99, 96. G Quantitation of percentage of hubs that show either average interior enrichment index >2 or average surface enrichment index >1.1. n = 120, 202. H Representative fluorescence images of cells containing nanobody coacervate hubs showing two topologies of GFP recruitment: surface or interior; higher in the +CQ case. Coacervates formed from 0.5 mg/mL HBpep-SA co-assembled with 0.1 mg/mL VHHGFP4-mCherry were added to the cell media with or without 100 µM Chloroquine. CQ refers to Chloroquine. Scale bar: 10 µm. Source data are provided as a Source Data file.
