![Figure 5. P4M binds endosomes/lysosomes. (A) COS-7 cells expressing mCherry (mCh)-tagged P4M were loaded with 100 nM LysoTracker green. The insets 1 and 2 are shown as a montage of images acquired at 3.9-s intervals showing motile, P4M, and LysoTracker-positive structures highlighted by the arrowheads. (B) COS-7 cells transfected with GFP-P4M, the late endosomal marker Rab7 (tagged with mTurquoise2 [mTq2]), and the early endosomal marker Rab5 (tagged with mCherry). A binary mask derived from the Rab7 and Rab5 images was used to calculate the relative intensity of the P4M signal associated with each compartment. The results from 24 cells are shown as a paired scatter plot. (C) As in B, except cells were transfected with the GFP-tagged FYVE domain from EEA1 as a marker of PtdIns3P. Bars: (main images) 10 µm; (insets) 5 µm. Statistics report the p-value from a paired Student’s t test (see Materials and methods).](https://cdn.rupress.org/rup/content_public/journal/jcb/205/1/10.1083_jcb.201312072/6/jcb_201312072_fig5.jpeg?Expires=1773089505&Signature=NpXXuTRo8RTrhvRu2sDxTnkaosSC~my-kSRjwnOt9wsnhAJWWuLUEaRZ36jl4HPlxSqX6cx85RT-lduJZGM9s~CmyBGy5MT21hYJKdjVxgPf-uvaiySViXaJNwuVzd9gKQQ6g6MnAghqVmVv3zTJGwXg0wPaFbt0QrvrtsHbLyl5iCdY3ugV-fZZNgjP2WdaQ~kQJVCYU6MYW6gLS96EgpLYBERFh4YY5LlUYW3DqJ5XD9xboFp40yNZJEmOk8Gfz2UF1aaiwUlZTBoNvIRdc5zhlOn5DWoDP2HBhadtx-nGPMPBRzsbpdQ6kR~hi2WOmjt8a5YZ~hwvQbiXq~DThQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
P4M binds endosomes/lysosomes. (A) COS-7 cells expressing mCherry (mCh)-tagged P4M were loaded with 100 nM LysoTracker green. The insets 1 and 2 are shown as a montage of images acquired at 3.9-s intervals showing motile, P4M, and LysoTracker-positive structures highlighted by the arrowheads. (B) COS-7 cells transfected with GFP-P4M, the late endosomal marker Rab7 (tagged with mTurquoise2 [mTq2]), and the early endosomal marker Rab5 (tagged with mCherry). A binary mask derived from the Rab7 and Rab5 images was used to calculate the relative intensity of the P4M signal associated with each compartment. The results from 24 cells are shown as a paired scatter plot. (C) As in B, except cells were transfected with the GFP-tagged FYVE domain from EEA1 as a marker of PtdIns3P. Bars: (main images) 10 µm; (insets) 5 µm. Statistics report the p-value from a paired Student’s t test (see Materials and methods).
