Fig 4.

The ATPase domain of Hsc70 is not required for its interaction with μNS. (A) Diagram of Hsc70 subdomains and truncation mutants. SBD, substrate-binding domain. (B) 293F cells were cotransfected with the μNS-encoding plasmid and either the HA-tagged ATPase domain or the HA-tagged SBD of Hsc70. HA-tagged constructs were immunoprecipitated from cell lysates with an anti-HA MAb and were then immunoblotted (IB) and probed for the HA tag or μNS. A control antibody (anti-Tsp) did not immunoprecipitate tagged Hsc70 or μNS. (C) 293F cells were cotransfected with plasmids encoding μNS and the HA-tagged truncation mutants of Hsc70 diagramed in panel A. Immunoprecipitates were prepared and immunoblotted as for panel B. (D) IF images of CV-1 cells ectopically expressing μNS and Hsc70(1-394) (top) or μNS and Hsc70(395-540) (bottom). Cells were fixed with 100% methanol. Bars, 2 μm.