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ARÁNZAZU MEDIERO Y COLS. AN. R. ACAD. NAC. FARM.

both at 12 and 24 hours after wounding (Figure 2A and 2B). 36
hours after the wounds were performed, P2Y2 staining was similar
in the three different treatments, including the siRNA treated
corneas, revealing a full recovery of the P2Y2 receptor (Figure 2C).

These results indicate that silencing the P2Y2 receptor in our
model was detected 12 hours after the wound was performed.
Nevertheless, P2Y2 receptor signal was again visible 36 hours after
the wound had been made.

Figure 1. P2Y2 receptor location in the cornea. Immunocytochemical analysis
for the P2Y2 in the cornea revealing the presence of the P2Y2 receptor in the corneal
epithelium (green fluorescence by FITC). Image managed with the Pascal software
of the Axiovert 200M confocal microscope at 40X magnification.

3.3. Over-expression of P2Y2 receptor after silencing

In half of the siRNA treated corneas, and 36 hours after the
wounding (96 hours after the first siRNA instillation), we have
observed an increase in the expression of P2Y2 receptors compared
with control and Ap4A (Figure 3). In this case, the P2Y2 signal was
not constrained to the outer layer of the epithelium, and it was
possible to localize P2Y2 receptors in the whole epithelium and in
the stroma (Figure 3).

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