Figure 2

Screen quality control and normalization. (A) Overview of intoxication assay represented in the demonstration dataset. HeLa cells stably expressing a short half-life luciferase (Luc2CP) were treated with PE or Ricin toxins. After binding at the cell surface, both toxins are internalized into endosomes and transported to the Golgi and then to the ER, where they translocate to the cytosol. Both toxins then inhibit translation and luciferase production. The three highlighted genes, CLTC1, STX16, and KDELR1, act on endocytosis, transfer to the Golgi, and Golgi to ER traffic, respectively. (B) Workflow of the genome-wide screen: Cells were seeded in 384-well plates pre-printed with siRNA for reverse transfection and incubated for 72 hr, then challenged with PE or Ricin for 8 hr before luciferase signals were measured. (C,D) Scatter Plot, with controls highlighted (C), and Box Plot of controls (D) of Replicate 1 of raw PE luciferase signal using the Quick Analysis module. (E) Plot of Replicate 1 versus 2 of raw PE luciferase signals. (F) Scatter Plot of Replicate 1 of raw PE luciferase signal with STX16 positive control highlighted. (G) Average Z-score-normalized PE luciferase signals with STX16 highlighted. (H) Average control (STX16)-normalized PE luciferase signals with STX16 highlighted.