Figure 1.
Global RNAi screen of Salmonella-infected human epithelial cells.
(A) Procedural outline for the screen. (B) The plate layout. Wells that contained siRNAs that result in cell death are in yellow and served as a positive control for transfection. The wells colored red contained anti-AKT1 siRNA, which served as a positive control for the reduction of intracellular S. typhimurium growth. The wells colored black contained non-specific siRNA and the wells colored green contained anti-GFP siRNA, which is unrelated to the GFP expressed by the bacteria. Both the non-specific siRNA and anti-GFP siRNA served as negative controls. Water wells, intended to reduce edge effects are in grey, and the experimental wells are in blue and purple.
Figure 2.
Four prominent sub-networks and associated functions that could plausibly play roles in promoting microbial growth.
(A) Cellular development, cellular growth. (B) Cell death. (C) Cell cycle. (D) Carbohydrate metabolism. The shaded molecules are the ones identified in the screen. The others were added by IPA to generate the sub-networks.
Figure 3.
The over-represented molecular and cellular function categories of the entire up and down networks.
The entire IPA-generated networks that modulate the intracellular growth of S. typhimurium were analyzed for over-represented functional categories. (A) The network that promotes growth. (B). The network that restricts growth. The significance threshold (p = 0.05) is the red, dashed line. The X axis is the –log of the p-values.
Table 1.
Some of the more interesting hits identified and their characteristics.
Figure 4.
Raw microscopy data from the screen with some of the more interesting hits.
Host cells are in blue, S. typhimurium is green. Only one representative non-silencing control (upper left corner) and one positive control (anti-AKT1), which is adjacent to the non-silencing control are shown due to space constraints. All of the positive and negative controls appeared similar.