Fig. 4: TpoR preferentially traffics through autophagosome-lysosome in the presence of JAK2 V617F. | Leukemia

Fig. 4: TpoR preferentially traffics through autophagosome-lysosome in the presence of JAK2 V617F.

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

Fig. 4

A The subcellular localization of TpoR was analyzed by confocal microscopy. DU145 cells expressing TpoR along with JAK2 WT, JAK2 V617F or Atg5 and stained for organelle marker (green; LAMP1) and antibodies against HA tag for TpoR (red). The scale bar represents 10 μm. Arrowheads indicate colocalization of TpoR in LAMP1 lysosomes. B The signal intensities of TpoR puncta and LAMP1 or LC3 puncta in (A) were calculated as gray values and the percentage colocalization of TpoR was calculated in each case. Data represents percentage of colocalization ±SEM. Error bars represent SEM. p-value is indicated in each graph. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. No. of cells n = 5–10 for each case, no. of vesicles n = 556–608 for LAMP1. C Cell surface expression of TpoR was investigated by flow cytometry using CD110-APC antibody against surface TpoR. Median fluorescence intensity (MFI) is indicated in each case. Representative image of the flow analysis is shown (n = 3).

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