Fig. 6. Dynein colocalizes with nascent inclusions by 5 hours postinfection and remains associated with the inclusion as late as 24 hours postinfection. Cos-7 cells were infected with C. trachomatis for 5 or 24 hours in the presence or absence of nocodazole. Cells were stained for simultaneous visualization with mAb DIC 70.1 to visualize the dynein intermediate chain and a polyclonal antibody to L2 EBs to visualize the nascent inclusions. By 5 hours postinfection, the dynein antibody revealed staining consistent with it being localized along microtubules, as well as associated with the aggregated chlamydiae. In cells treated with nocodazole, dynein staining of the microtubules is absent and much of the staining is evident around the nuclear envelope. The nascent inclusions show dramatic dynein recruitment even in the presence of nocodazole. Dynein remains associated with the mature chlamydial inclusion even after infection for 24 hours. Bars, 10 µM.