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Fig. 1. Wnt gene expression in the mouse lens and activation of Wnt signaling by
vitreous humor. Total RNA was isolated from E16, P4, P21 and adult rat lenses
(A), or from rat (P4) lens central epithelium (CE) and equatorial epithelium,
which contains both the proliferating cells and elongating cells (EC) (B).
cDNA was prepared by reverse transcription and amplified with primers specific
for each Wnt. Each reaction was normalized to ß-actin. Analyses of at
least three different RNA preparations from the same tissues provided similar
results. (C) Mouse lens cells were transiently transfected with TOPFLASH or
FOPFLASH reporter plasmid, and then stimulated with vehicle (lanes 1, 7),
aqueous humor (lane 2), vitreous humor (lane 3), control medium (lane 4),
5xWnt3a conditioned medium (lane 5), 10xWnt3a CM (lane 6), 1 ng/ml
EGF (lane 8), 50 ng/ml FGF2 (lane 9), 10 ng/ml PDGF (lane 10), or 5 ng/ml
TGF-ß(lane 11) for 16 hours. Cell lysates were assayed for luciferase
activity. (D) Control medium (lane 1), vitreous humor pre-incubated with
control medium (lane 2), vitreous humor pre-incubated with sFRP-1-conditioned
medium (lane 3), or sFRP-1-conditioned medium alone (lane 4) were added to
mouse lens cells transfected with the TOPFLASH reporter plasmid. Cell lysates
were analyzed by luciferase assay.
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