Fig. 6: Co-localization of Y. pestis ΔyspE2 with hGBP1 dramatically promotes the clearance of bacteria during infection of HeLa cells.
From: Subversion of GBP-mediated host defense by E3 ligases acquired during Yersinia pestis evolution
a HeLa cells stably expressing GFP-hGBP1 were infected with the RFP-expressing wild type Y. pestis 201, ΔyspE2, ΔyspE2 complemented with the plasmids expressing YspE2 or YspE2C386A at an MOI of 10. After 1 h of infection, the confocal micrographs were acquired using an Andor Dragonfly spinning disk confocal microscope. The far right column showed the 3D reconstruction images from the multiple slice images of the enlarged framed areas. The intracellular bacteria of Y. pestis ΔyspE2, but not 201, was significantly colocalized with hGBP1. b Representative frames from Supplementary movie 1 to 4 of HeLa cells expressing GFP-hGBP1 infected with wild type, ΔyspE2, ΔyspE2/YspE2 or ΔyspE2/YspE2C386A RFP-expressing Y. pestis. HeLa cells were seeded in Lab-Tek chambered cover-glass and infected with the indicated Y. pestis strains and the images were taken every 5 mins until 9 h of infection using spinning disk system with Nikon ECLIPSE Ti-E microscopy. These experiments have been repeated at least for three independent times with similar results.
