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Figure 2 | Journal of Molecular Signaling

Figure 2

From: Heterodimerization of β2 adrenergic receptor and somatostatin receptor 5: Implications in modulation of signaling pathway

Figure 2

Microscopic Pb-FRET analysis in HEK-293 cells coexpressing cMyc-β 2 AR/HA-hSSTR5. (A) Representative photomicrographs of HEK-293 cells illustrating cMyc-β2AR (red), HA-hSSTR5 (green) and colocalization in merged image (yellow) (Panel a). Pb-FRET microscopy on HEK-293 cells coexpressing cMyc-β2AR/HA-hSSTR5 was performed as described in Material and Methods. A selection of photomicrographs illustrating photobleaching profile of donor alone and in the presence of acceptor are shown in panels b and d. Histograms shown in panels c and e represents pixel by pixel analysis of time constant of donor and donor + acceptor and the mean time constant shown in black was calculated from a Gaussian distribution curve. Note the increased time constant of donor in presence of acceptor, indicating interactions between cMyc-β2AR/HA-hSSTR5. (B) HEK-293 cells expressing cMyc-β2AR/HA-hSSTR5 were treated with SST (1 μM) and formoterol (1 μM) in combination for 15 min at 37°C. Activation of hSSTR5 with SST and β2AR with formoterol simultaneously displayed increase in the effective FRET efficiency. Representative photomicrographs illustrating bleaching profile of the donor in the absence or presence of acceptor are shown in panels b and d. Histograms shown in panels c and e represent pixel by pixel analysis of time constant of the donor alone or donor + acceptor respectively in cells treated with SST and β2AR specific agonist together for 15 min. Data are representative of three independent experiments performed in duplicate. Approximately 60-70 cells were analyzed per experiment.

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