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. 2016 Dec 21;12(12):e1006101. doi: 10.1371/journal.ppat.1006101

Fig 1. Interaction between Cig57 and FCHO2.

Fig 1

(A) A yeast-two hybrid screen, using a HeLa cDNA library, revealed FCHO2 as a binding partner of Cig57. Growth on double dropout (DDO) plates for all transformations indicates plasmid retention and yeast viability. Growth on quadruple dropout (QDO) plates indicates the interaction between FCHO2 and Cig57 (segment 2) and the lack of growth in segment 1 and 3 of this plate indicates no interaction with the empty vectors present. (B) A Pull-down assay verified the interaction between Cig57 and GFP-FCHO2 during Coxiella infection of HEK 293T cells. Cells were infected with wild-type (WT) Coxiella, or Coxiella cig57::Tn strain expressing 3xFLAG-Cig57 from a plasmid, and transfected with GFP constructs (GFP or GFP-FCHO2). GFP proteins were pulled out using GFP-Trap beads. The presence of 3xFLAG-Cig57 in the pull down lysates indicates interaction between GFP-FCHO2 and Cig57. Results are shown for one repeat of the experiment. (C) Representative images showing HeLa cells transfected to express mCherry or mCherry-Cig57 (red) during infection with wild-type C. burnetii (green). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (D) HeLa cells stably expressing GFP-FCHO2 were transfected with pmCherry-Cig57 and nuclei stained with DAPI (blue). Scale bars represent 10 μm and the CCV is denoted by an asterisk.