Development of a HCI-based assay to assess phenotypic changes to the Golgi.

<p>(<b>A</b>) IFA showing co-localization of RVFV Gn (Red) and a GFP-fused Golgi marker (Green) in a HeLa-G cell line that stably expressed RVFV Gn protein. (<b>B</b>) Software development for HCI-based analysis of Golgi. Acquired confocal images of nucleus (I), cell (II) and the Golgi complex (III) in mock-treated HeLa cells. The juxtanuclear Golgi is qualified as “intact Golgi” by image analysis and is marked by the red boundary around the clustered Golgi stacks (insert, IV) in mock treated cells. In contrast, image analysis disqualified many of the numerous scattered spots in the cytoplasm of D011-2120-treated cells as “intact Golgi” (insert, V). (<b>C</b>) Heat map of Golgi number and Golgi area by HCI-based analysis with increasing concentrations of the hit compounds and reference compounds. (<b>D</b>) Dose response curve analysis of lead compounds showing an increased reduction in Golgi as a function of compound concentration. Percent Golgi number is shown in blue and percent infection inhibition is shown in green.