TMEM16F resides in late but not early or recycling endosomes. (A–D) Jurkat cells expressing TMEM16F-RFP together with Rab7-GFP (A and D), Rab5-GFP (B), and Rab11-GFP (C) were stimulated on coverslips coated with αCD45 (D) or αCD3 (A–D). Cells were imaged by TIRF (A–C) or confocal microscopy (D). (A–C) Spatial correlation of localization of TMEM16F and Rab7 (A), Rab5 (B), or Rab11 (C) upon TCR stimulation. The fluorescence intensities (pixels) along the dotted line were determined by ImageJ (right). Images are representative of six to seven cells. Bars, 5 µm.