Figure 3: Recycling of unliganded EphA2 through the RE. | Nature Communications

Figure 3: Recycling of unliganded EphA2 through the RE.

From: Ubiquitination switches EphA2 vesicular traffic from a continuous safeguard to a finite signalling mode

Figure 3

(a) Colocalization of EphA2 and Rab11a. Left column: immunostaining of endogenous EphA2 (upper) and Rab11a (middle) in fixed Cos-7 cells, green/magenta overlay (lower). Right column: fluorescence images of ectopically expressed EphA2-mCitrine (first) and bfp–Rab11a (second) in living Cos-7 cells, green/magenta overlay (third). Fourth row represents magnified insets from respective overlay ROIs (yellow boxes). (b) Fluorescence redistribution after photoactivation of EphA2–paGFP in Cos-7 cells. First row: EphA2–paGFP fluorescence at the indicated times (s) after photoactivation on the RE, second: EphA2-mCherry fluorescence, third: bfp–Rab11a fluorescence. Lower graphs: ratiometrically determined decrease of EphA2–paGFP/EphA2-mCherry fluorescence at the RE (left) and gain at the PM (right). An average of 14 independent fluorescence loss and recovery curves (grey) were fitted to an exponential function to retrieve half-times (t1/2). Scale bars: 10 μm.

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