Fig 5.TIF (2.31 MB)

Dynamic and lifetime analysis of the GlCHC:GFP reporter.

figure

posted on 2016-07-20, 03:30 authored by Jon Paulin Zumthor, Lenka Cernikova, Samuel Rout, Andres Kaech, Carmen Faso, Adrian B. Hehl

(A-D) FRAP and iFRAP analyses were done in the same field of view including an unbleached control cell (ROI05). FRAP analysis (ROI02, ROI03) shows no recovery whilst iFRAP analysis (ROI04) shows no loss of fluorescence after 225s, indicating no measurable turnover for the GlCHC-GFP reporter. Pre-bleach (B), post-bleach t0 (C), post-bleach t220 (D). (E-H) Lifetime analysis of individual GlCHC assemblies over 30min. The insets F and G show precise recognition of individual fluorescent events by the spot tracking tool in IMARIS. Clathrin assemblies are labelled in green whereas individual tracking spots are labelled in purple. (H) Analysis of 187 individual fluorescent events revealed lifetimes between 30-180s (9.65%), 210-900s (56.15) and 930-1800s (34.20%).