Fig. 4.

EspG has no effect on the uncoating of TfR/EEA1 positive vesicles

A) HeLa cells expressing mRFP-CLC were infected with EHEC WT, a ΔespG mutant or ΔespG + pEspG:4xHA. After 5 h of infection cells were fixed and labeled for EEA1 and TfR. During WT EHEC infection, enlarged TfR positive, EEA1/CLC negative structures can be seen in infected cells. These structures were not observed in non-infected cells, nor those infected with EHEC ∆espG, but were more pronounced in cells infected with EHEC ΔespG + pEspG:4xHA. Scale bars represent 10 μm, white arrow heads indicate triple-positive structures, yellow arrow heads indicate large TfR-positive structures lacking EEA1/CLC. B) Quantification of images shows that EspG has no effect on the association of clathrin with TfR positive early endosomes (n = 2, > 150 EEA1/TfR positive structures scored per condition per experiment, graph is representative of one independent experiment, ns = non-significant).