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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.