Figure 6.
PIKfyve inhibition doesn’t affect the endocytosis of PFFs. (A–I) iNs were incubated with (A and B) 20 µg/ml Transferrin AF647 for 5 min, (C and D) 20 µg/ml Dextran-AF647 for 2 h, or (E and F) 4 µg/ml PFF-AF647 for 3 days, in the absence (A, D, and G) or presence (B, E, and H) of 100 nM Apilimod. Cells were imaged using live-cell 3D spinning-disc confocal microscopy. Maximum z-projection images are representative of three biological replicates for A and B and four biological replicates for D–H. Cells are outlined with dotted lines. Scale bar: 10 µm. Insets show a 3× magnification with a scale bar of 3.3 µm. (C, F, and I) Quantitative analysis shows that 100 nM Apilimod did not affect the uptake of Transferrin, Dextran-AF647, or PFF-AF647. Each dot represents the total relative fluorescence intensity per cell for each internalized cargo. The number of cells analyzed is indicated.

PIKfyve inhibition doesn’t affect the endocytosis of PFFs. (A–I) iNs were incubated with (A and B) 20 µg/ml Transferrin AF647 for 5 min, (C and D) 20 µg/ml Dextran-AF647 for 2 h, or (E and F) 4 µg/ml PFF-AF647 for 3 days, in the absence (A, D, and G) or presence (B, E, and H) of 100 nM Apilimod. Cells were imaged using live-cell 3D spinning-disc confocal microscopy. Maximum z-projection images are representative of three biological replicates for A and B and four biological replicates for D–H. Cells are outlined with dotted lines. Scale bar: 10 µm. Insets show a 3× magnification with a scale bar of 3.3 µm. (C, F, and I) Quantitative analysis shows that 100 nM Apilimod did not affect the uptake of Transferrin, Dextran-AF647, or PFF-AF647. Each dot represents the total relative fluorescence intensity per cell for each internalized cargo. The number of cells analyzed is indicated.

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