Figure 4

Suppressive activities measured with fluorescent reporters. Suppressive activities were screened using gene-specific fluorescent fusion reporters in a transient expression assay. The 96 hairpins were individually transfected with a target-specific GFP fusion reporter, and an AsRed-1 non-specific reporter. Specific activity is shown as percentage fluorescence of the unsuppressed control (black bars above; control shown in blue), and non-specific activity is shown as the fold difference (normalization factor) relative to the baseline non-specific activity of the unsuppressed control (red bars below, control shown in blue). E.g. a normalization factor of 1 is no non-specific activity, a value of 2 is twice as much non-specific activity of the baseline control (set at 1). Categorical markers divide the hairpins into inactive, active and highly active groups. The reporter used for each hairpin (green bars), and the 22 targeted regions (brown bars) are also shown. (a) Suppressive activities measured with the first round 'long' (core genes) and 'short' (accessory genes) reporters. (b) The previous long reporters were replaced with a series of shorter fragment reporters and suppressive activities were remeasured. The suppressive activities form the previous short accessory gene reporters are included for comparison (hollow bars).