Figure 2.

Ectopic expression of wildtype Shep E but not Shep E^RRM mutant decreased gypsy barrier activity in muscle. A. Anterior thirds of wildtype larvae expressing no transgene, UAS-shep E, or UAS-shep E^RRM driven by arm::Gal4 were used for western blotting for Shep E and a loading control Protein on ecdysone puffs (Pep). Normalized band intensity of Shep E relative to Pep was quantified by CCD imaging of chemiluminescence followed by Photoshop analysis. B. Schematic diagram of UAS-luciferase system shows flanking gypsy insulator sequences act as a barrier to allow luciferase expression. C. Relative luciferase activity of non-insulated (left) or insulated (right) reporters in individual larvae expressing UAS-su(Hw) RNAi, UAS-shep E, or UAS-shep E^RRM driven by Mef2::Gal4. Luciferase activities are reported as boxplots with boxes representing the first and third quartiles. Luciferase activities across genotypes were compared by One-way ANOVA followed by Tukey HSD post hoc tests, with statistical threshold at 0.05. For each genotype, luciferase signals were read for 12 individuals, each normalized to input protein level. Samples showing signal variation more than 100-fold from median of all replicates were discarded as outliers. At least 10 valid replicate samples of each genotype were used for statistics.