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Designing the siRNA library screen.

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posted on 14.10.2015, 04:05 by Maja K. Choma, Jennifer Lumb, Patrycja Kozik, Margaret S. Robinson

The screen was carried out on four sets of standard plates and four sets of plates that were “sensitised” by including siRNA targeting the AP-2 μ2 subunit in every well. The same positive and negative control siRNAs were added to the first two columns of every plate. Cells were added to the 8 sets of plates on Day 1; then on Day 2, half of the plates (2 standard and 2 sensitised) were treated with 4-HT and the other half with ethanol as a control. On Day 3, the plates were shifted to a lower temperature to enhance Nef activity, and on Day 4 the assays were carried out.