Fig. 4: Reversible and dynamic protein shuttling in living cells.
From: Mandipropamid as a chemical inducer of proximity for in vivo applications
a, Schematic illustration of the four-step procedure to shuttle cytosolic protein between different intracellular targets. b, Confocal fluorescence microscopy images of the shuttling process between vimentin and mitochondria in a living cell. COS-7 cells were cotransfected with vimentin-mNeonGreen-PYRMandi-IRES-Halo-ABI and TOM20-SNAPf-PYL. Halo–ABI and SNAPf–PYL were labeled with HTL-SiR and tetramethylrhodamine (TMR)-Star, respectively. The top row shows dynamic receiver localization, and the middle row shows receptor localizations as references. Split images depict vimentin and mitochondrial localization in two different channels. The bottom row shows respective merges. Images were acquired at the indicated times before and after the addition of ABA-AM (200 nM), after the addition of revABA (20 μM) and after the addition of Mandi (200 nM); scale bar 20 µm. Data are representative of 22 cells from two independent experiments. c, Pearson correlation coefficients (PCC; mean ± s.d.) between receiver and respective receptor channel images at the indicated time points for four-step shuttling between cytosol, mitochondria and vimentin as shown in b. Small symbols represent individual cells at the indicated time points. In the inset, conditions were compared with a two-sided paired t-test.
