Fig 3.

CpoS deficiency disrupts the formation of inclusion membrane microdomains. (A) Co-IP confirms the interaction between CpoS and IPAM. HeLa cells were co-infected with CTL2-cpoS::cat/pCpoS(L2)-FLAG (2.5 IFU/cell) and CTL2/pIPAM(L2)-MYC, CTL2/pIncA(L2)-MYC, or CTL2/pIncD(L2)-MYC (2.5 IFU/cell), or were infected with each strain individually (5 IFU/cell). CpoS-FLAG was precipitated from lysates at 26 hpi. Samples were analyzed by western blot analysis (* indicates visible Co-IP of MYC-tagged Inc). (B) Enrichment of CpoS-FLAG in IPAM-containing microdomains in HeLa cells infected with CTL2-cpoS::cat/pCpoS(L2)-FLAG (2.5 IFU/cell, 24 hpi, scale = 40 µm). (C–F) Localization of IPAM (C), IncB (D), CTL0475 (E), and active Src kinases (F) in infected HeLa cells (5 IFU/cell, 20–24 hpi, scale = 20 µm, confocal). The proteins were detected by immunostaining of endogenous proteins (C, E, and F) or by using the split-GFP approach (D). (G) Architecture of the microtubule cytoskeleton in infected HeLa cells (5 IFU/cell, 22 hpi, scale = 20 µm).