Co-visualization of Hok11–624 and EEs in hyphal cells of the mutant Δhok1. Hok11–624 was fused to GFP (green) and coexpressed with mCherry-Rab5a (red). The Hok11–624 protein is missing the C-terminal region and therefore is not able to interact with the endosomes. However, it is still moving in retrograde direction, suggesting that it binds to dynein. The subapical regions were photobleached to avoid signal interference. This treatment did not harm retrograde motor or EE motility (Schuster et al., 2011b). Cells were placed on an agar cushion, and image series were taken immediately using a charge-coupled device camera (CoolSNAP HQ2; Photometrics) and a laser-based inverted microscope (IX81 [Olympus] equipped with a VS-LMS4 Laser Merge System [Visitron Systems] and controlled by the software MetaMorph). Frames were taken every 150 ms for 11.1 s. Images were adjusted in brightness, contrast, and γ settings. Time in seconds and milliseconds; bar is in micrometers.
