Critical Role of FLRT1 Phosphorylation in the Interdependent Regulation of FLRT1 Function and FGF Receptor Signalling
Figure 5
Differential trafficking and localization of activated FGFR1 by FLRT1 and Y3F-FLRT1.
A–F) Representative confocal microscope images of SH-SY5Y cells grown to confluency on coverslips and co-transfected with FGFR1, and either FLRT1-HA (A–C) or Y3F-FLRT1-HA (D–F) Cells were fixed with either no stimulation, A) and D), or after exposure to FGF2 (20ng/ml) in the presence of heparin (10mg/ml) for 5 min, B) and E), or 30 min, C) and F) Cells were stained with anti-pY766FGFR1 (pFGFR1, red), anti-HA (FLRT1 and Y3F-FLRT1, green) and anti-pY416Src (pSrc, blue) Merged images (and inset magnified images of cell body and neurites) demonstrated co-localization of FLRT1/FGFR1 (yellow), FLRT1/pSrc (cyan), pFGFR1/pSrc (magenta) and FLRT1/pFGFR1/pSrc (white, white arrows) Images were acquired on a Leica confocal microscope and processed using Image J and Adobe Photoshop 60 and represent at least 3 independent experiments• Scale bar: 20mm.