Extended Data Figure 4. Formation of PS⁺ structures during Lm infection.

Low magnification images used to generate images shown in Figure 2a. HeLa cells were infected with wild type Lm expressing RFP for 6 h and then cooled on ice and stained with a fluorescent probe (Annexin V-Alexa 488) for 10 min to label exofacial PS. Cells were then fixed and analyzed by fluorescence microscopy to identify PS⁺ structures and bacteria. Scanning electron microscopy (SEM) of the same cell revealed that PS⁺ structures were associated with the dorsal surface of infected cells. Differential interference contrast (DIC) microscopy of cells was also performed to help identify cells for correlative imaging analysis. Images representative of 2 independent experiments. Scale bars, 20 μm.