Figure 4
From: H-Ras transfers from B to T cells via tunneling nanotubes
Recovery dynamics of GFP-H-RasG12V differ according to its cellular localization. B721.221 cells expressing GFP-H-RasG12V and PKH26GL red-labeled Jurkat cells were seeded and imaged as detailed in Figure 2. GFP-H-RasG12V was bleached at three different cellular localizations: the ventral membrane of the donor B721.221 cell; the central portion of a B721.221/Jurkat cell-connecting TNT and a GFP-H-Ras-G12V-labeled patch on a Jurkat cell bordering a TNT originating from a B721.221 transfectant. (a) Panels depict typical confocal micrographs prior to bleaching, immediately post-bleaching and after a 2-min period of recovery at each of the cellular localizations. Scale bars represent 1 μm. (b) Initial fluorescence intensities in FRAP measurements were determined as 100%. Graph depicts the mean±S.E.M. of the normalized fluorescence of 10 independent measurements for each of the cellular localizations. At the indicated section of curve b and throughout the entire measurement of curve c, the relative fluorescence intensities were significantly different than the analogous sections of curve a, by two-tailed Student’s t-test (*P<0.05, ***P<0.001)
