Figure 1.

NURF301 affects the formation of gypsy insulator bodies. (A) Localization of insulator bodies is disrupted after depletion of individual gypsy insulator proteins. Right panels are quantification of average total GFP foci number per cell and average number of large foci (volume > 0.5 μm³) per cell imaged on glass slides. Cells were grown at 25°C. (B) Workflow of high-content RNAi imaging screen: Pre-spot dsRNA on plates, dispense cell suspension, incubate 4 days at RT then 2.5 h at 4°C, fix and DAPI stain, image, and process data. (C) Average large foci number per nucleus after each dsRNA treatment. Z’ factor = 0.8, indicating a good assay. (D) Verification on glass slides that NURF301 knockdown can alter the formation of gypsy insulator bodies. Cells were grown at 25°C, and these samples were processed in parallel with cells in (A). Quantification of the average total foci and large foci number are shown on the right. Scale bars: 5 μm. Comparisons in groups are all compared with mcherry dsRNA knockdown control cells. Three biological replicates were performed with n > 500 cells examined in each condition. Two-tailed unpaired t-test was used, and error bars show standard deviation. **P < 0.01 and ^****P < 0.0001.