Extended Data Fig. 3: TME biocompatibility and its functionality to enrich unfolded proteins in vitro and cellular proteins from cell lysate via RUBICON workflow.
From: Global analysis of endogenous protein disorder in cells
a, Cell viability assays for cells stained with 25 – 75 µM TME for 30 and 60 min. Cells were also incubated with the same amount of DMSO as vehicle controls. n = 4 biological replicates; mean ± s.d. b, Confocal microscopy images of cells stained with TME and co-stained with ER-Tracker and DRAQ5 for endoplasmic reticulum (ER) and nucleus respectively. Scale bar: 20 µm. c-d, Only TME labeled model proteins (c) and proteins from cell lysates (d) were enriched by the RUBICON workflow, which displayed evident fluorescence in TME, SYPRO Ruby and Coomassie Blue channels. The input loading amount for BLG and cell lysate was 7.5 and 50 µg respectively. SYPRO Ruby were used as a sensitive stain to visualize all proteins while Coomassie Blue were used to quantify total protein amount. “TME-/BLG-” and “TME-/Lysate-” denote samples that contained beads only. e, Flow cytometry results showing that proteostatic stressor (that is, tunicamycin and MG132) treated cells displayed significantly higher TME fluorescence compared to control. Gating strategies are shown in Supplementary Fig. 6. n = 3 biological replicates; mean ± s.d.
