Figure 6. Dimerization at individual mitochondria, centrosomes and kinetochores.

Cells expressing mCherry–eDHFR and Haloenzyme–GFP-anchor domain fusion proteins specific for (a) mitochondria (ActA), (b) centrosomes (AKAP9) or (c) kinetochores (Nuf2) were incubated with 20 µM cTMP-Htag for 1 h, then washed before imaging. Cells were imaged before and after targeted laser illumination, as indicated. Individual structures in these cells (indicated by arrowheads in insets) were targeted with a 10–100ms pulse from a 405nm laser immediately before the ‘post’ image. Insets show boxed regions in GFP (top row) and mCherry (middle row) and colour-merge (bottom row) from indicated time points. GFP is locally photobleached by the 405 nm uncaging pulse. Gaussian smoothing with a radius of 1 pixel was applied to mCherry images in c. Scale bars, 5 or 1 µm in insets.