Fig. 1. Sequestration of Xena after the expression of mitochondria-binding Ena/VASP proteins and its effects on growth cone morphology. (A-C) FP4-Mito-GFP (green, A) colocalized with the mitotracker (red, B) in retinal axons (arrowheads, C). (D-F) Xena-GFP (green, D) colocalized with FP4-Mito-RFP (red, E) in retinal cells co-expressing both constructs (F) in cryostat sections of stage 42 eyes. Arrowheads show the regions in the inserts. Inserts highlight the sequestration of Xena by the Mito construct. (G,H) In a stage 39 retinal explant, Xena-GFP was enriched along the retinal axon shaft and at the tips of growth cone filopodia (arrows). (H) Phase-contrast picture of G. (I) Phase-contrast picture of J-L. (J-L) Expression of the FP4-Mito construct (red, K) selectively depleted Xena-GFP (green, J) from sites of normal localization and sequestered it on the mitochondrial surface (L). (K) RFP-labeled mitochondria in a retinal axon expressing FP4-Mito-RFP. (M-O) Growth cone morphology of control GAP-GFP-labeled (M), control RFP-labeled (N) or FP4-Mito-GFP+RFP-labeled inactivated (O) retinal ganglion cells (RGCs) in explant culture. RGC, retinal ganglion cells. Scale bar: 56 µm in M for D-F; 8.5 µm in M for A-C,G-O.