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Targeting sequences were chosen according to commercial software (si RNA Target Designer; Promega, Madison, WI).
The following sense and antisense sequences were used to target murine ICAM-1: sense, 5′-ACC GAT CGT CAC GGC GAT TTA TAA GTT CTC TAT AAA TCG CCG TGA CGA TCT TTT TC-3′; antisense, 5′-TGA AGA AAA AGA TCG TCA CGG CGA TTT ATA GAG AAC TTA TAA ATC GCC ACG AT-3′.
Two micrograms of basic vector-designed h MGFP vector (psi STRIKE; Promega; Fig.The si RNA are generated by an RNase III-like processing reaction from long ds RNA.The biggest challenge in the systemic use of si RNA-based therapies was delivering them to the cytoplasm.Green fluorescent protein (GFP) expression plasmid vector is used as a transfection marker in the retinal cells.ICAM-1 expression was analyzed by enzyme-linked immunosorbent assay and flow cytometry.
All experiments were performed in accordance with the guidelines of the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.