Fig. 1: pH-dependent ratiometric FRET of SRAI-TpoR. | Leukemia

Fig. 1: pH-dependent ratiometric FRET of SRAI-TpoR.

From: Autophagosome-lysosome mediated secretion of the thrombopoietin receptor is modulated by distinct driver mutations of myeloproliferative neoplasm

Fig. 1

A Schematic representation of SRAI-TpoR construct. B U2OS cells were transfected with JAK2 WT, JAK2 WT+ Bafilomycin A1, JAK2 V617F. Imaging of TOLLES-YPet signal was performed under a confocal microscope. (Left) Ratiometric FRET (Intensity of TOLLES/ Intensity of YPet) approach was used to generate the rainbow RGB images. The ratiometric FRET image (left panel) has been represented; overlay of the donor TOLLES signal (red) and FRET signal (green) has been represented (middle and right panels) to identify areas with (yellow) and without FRET (red). (center) Overlay of FRET signal with donor TOLLES signal; (right) Inset shows an enlarged view with arrowheads indicating puncta with quenched FRET signal. The scale bar represents 10 μm. C Imaging of TOLLES-YPet signal was performed at the cell surface. Overlay of FRET signal (green) with donor TOLLES signal (red) is shown. Arrowhead indicates quenched FRET signal. The scale bar represents 10 μm. D Data represents percentage of FRET puncta ± SEM. Error bars represent SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. No. of cells n = 10–12 for each case.

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