A High Precision Survey of the Molecular Dynamics of Mammalian Clathrin-Mediated Endocytosis (original) (raw)
Figure 5
Scaling relationships between CCS size and recruitment signatures.
(A–E) Analysis of cells coexpressing Clc-mCherry and TfR-phl. (A) Example scission event at a punctate CCS. (B) Example scission event at a larger CCS. The larger (brighter) CCS was associated with a larger (brighter) patch of TfR-phl at pH 7.4. (C) Histogram of normalised TfR7 fluorescence (_F_TfR7), averaged over −18 to −10 s preceding scission. Size classes defined as indicated: blue = {_F_TfR7 <33rd percentile}; green = {33rd percentile < _F_TfR7 <66th percentile}; red = {_F_TfR7 >66th percentile}. (Di) Average normalised _F_Clc signatures for classes defined in (C). The signatures are well separated, indicating that _F_Clc scaled with _F_TfR7. (Dii) Average normalised Clc-mCherry fluorescence signatures (_F_Clc7) for randomly allocated classes. Note the signatures are very similar. (Ei) Average normalised _F_TfR7 signatures for classes defined in (C). The signatures are well separated, as expected. (Eii) Average normalised _F_TfR5 signatures for classes defined in (C). The signatures are very similar, indicating that TfR5 fluorescence did not scale strongly with CCS size. (F) Example traces showing the scaling relationship between _F_TfR7 and the respective RFP recruitment signatures. (G) Stem plot of sum of differences between class averages and the overall average ordered by magnitude, colour coding as in (B). Grey bars indicate the 95% confidence interval for random event assignment to classes.