Figure 3: Loss of PHD3 inhibits EGFR internalization.
From: PHD3 regulates EGFR internalization and signalling in tumours
(a–d) EGFR internalization is inhibited on PHD3 loss. Immunofluorescence of G55 cells expressing control or PHD3 shRNA, non-stimulated or stimulated with EGF (20 ng ml−1) for the indicated times. EGFR co-localizes at early time points with EEA1, peaking at 15 min after stimulation (a,b). At later time points (30 and 60 min) after stimulation, EGFR co-localizes with the late endosomal marker mannose 6-phosphate receptor (M6PR; c,d). PHD3 loss compromises EGFR internalization, with EGFR remaining at the plasma membrane (a,c). The graphs show quantification of the increase in EGFR/EEA1 co-localization (b) and EGFR/M6PR co-localization (d) relative to non-stimulated cells (n=50–200). (e,f) Deletion of PHD3 blocks EGFR internalization. Internalization of EGFR in wild-type or PHD3−/− astrocytomas was quantitatively assessed by capture ELISA (n=6). The mean±s.e.m. is shown in the graph (e) and the table (f). All values are means+s.e.m., Student’s t-test *P<0.05; **P<0.01; ***P<0.001. Scale bars, 10 μm.
