Fig. 2
From: Intraoperative visualization of nerves using a myelin protein-zero specific fluorescent tracer
Localization of binding of P0 peptides to myelinating Schwannoma cells. Fluorescence confocal images of RT4 D6P2T Schwannoma cells after incubation with A C-terminally Cy5-labelled P01–25(Cy5-P01–25; I), P041–65 (Cy5-P041–65; II), P061–85 (Cy5-P061–85; III), P085–105(Cy5-P0185–105; IV), P095–120 (Cy5-P095–125; V) or P0101–125 (Cy5-P0101–125; VI). B Fluorescence confocal images of RT4 D6P2T Schwannoma cells after incubation with Cy5-P095–120 represented in 3D (I), and the extracellular portion of P0 (Cy5-P0ex; II), Cy5-labelled anti-P0 antibody clone H60 (Cy5-P0Ab-H60; III), a non-P0-specific peptide (Cy5-NP41; IV). Staining of non-P0-expressing MDAMB 468 cells with Cy5-P0101–125 (V), Cy5-P0Ab (VI) and Cy5-Maleimide (CI) were used to show specificity for P0. Blocking experiments showed a clear decrease in the mean fluorescence intensity for Cy5-P0101–125 after pre-incubation with P0Ab (II). Quantification of fluorescence intensity with and without blocking (III; Cy5-P0101–125 in red, blocked conditions in black) further underlined specificity (p = 0.001). In all confocal images, Cy5 is represented in red, nuclear staining (Hoechst) in blue and lysosomes (lysotracker green) in green