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Such a constant mean amount may be due to the formation of small clusters within early endosomes.
10.7554/eLife.06156.014 Figure 2. The constant mean amount of p-EGFR per endosome corresponds to receptor clusters that are regulated by Hrs and PTPN11.
Thus, the constant mean amount of p-EGFR in endosomes likely corresponds to the receptor clusters observed by super-resolution microscopy.
Figure 3C shows that this simple theoretical model can reproduce our observations of a constant mean amount of p-EGFR per endosome in a wide range of EGF concentrations when fitted to the experimental data.
To further validate that the constant mean amount of p-EGFR per endosome corresponds to receptor clusters, we performed a focused RNAi screen on established components of the endosomal receptor sorting machinery (CHLCb, CHLCa, Hip1, Hip1R, Htt, Tom1, Tollip, Tom1L1, Tom1L2, Hrs, Snf8, Vps24).
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By quantitative high-resolution FRET microscopy, we found that phosphorylated epidermal growth factor receptor (p-EGFR) is not randomly distributed but packaged at constant mean amounts in endosomes.
The coefficient β0 is a constant, representing the mean amount of fatigue in the control group at completion, given a score of zero fatigue at baseline.
We discovered that the mean amount of p-EGFR per endosome was fairly constant over time and p-EGFR was found in small clusters in early endosomes.
Strikingly, the mean amount of p-EGFR per endosome remained fairly constant, despite the EGF concentration varying almost over three orders of magnitude.
The coefficient β2 represents the mean amount of fatigue at completion amongst people who received acupuncture (the treatment), holding constant fatigue at baseline (β1).
with constant mean power.
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