Figure 2 | Scientific Reports

Figure 2

From: Vasopressin receptor 2 mutations in the nephrogenic syndrome of inappropriate antidiuresis show different mechanisms of constitutive activation for G protein coupled receptors

Figure 2

AVP-induced cellular redistribution of wild type V2R in cells expressing rGFP-tagged β-arrestin1 or β-arrestin2. Representative pictures are shown before (t = 0) and 60 min after application of 1 μM Arg-vasopressin (t = 60). Before dendra2 photo-conversion, the fluorescence signal in the green channel represents both receptor and arrestin proteins (dendra2 + rGFP). After photo-conversion (t = 0), green channel fluorescence mostly represents the distribution of β-arrestins (rGFP), whereas pure receptor signals (dendra2) appear in the red channels. Co-localizations of receptor and β-arrestins are seen as yellow signals in the superposed images. Experiments were carried out in KRH buffer at 37 °C. Note the formation of a punctuated perinuclear localization pattern of the receptor following the application of AVP (1μM). The apparent difference in fluorescence intensity between β-arrestin 1 and 2 in the picture (green channels after photo-conversion) does not reflect the difference in expression of the two isoforms, as the brightness of the pictures was adjusted independently in the two sets of cells. A microscopic quantification of β-arrestin 1 or 2 expressions in these cells lines is shown in Supplementary Fig. S3. Also, an approximate quantification of AVP-induced receptor internalization and β-arrestin colocalization is given in Supplementary Fig. S4.

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