FIG.5.

Expression of inPAK inhibits MV replication in permissive 3T3 cells. (A) Representative Western blots showing detection of activated, phosphorylated PAK-1 in uninfected 3T3.CD4 cells (lanes 1 and 4) and MV-infected (MOI of 5) 3T3.CD4 cells that were nontransfected (lanes 3 and 6) or transiently transfected with the PAK-1 autoinhibitory domain (inPAK, lane 2) or a noninhibitory PAK-1 domain (nonPAK, lane 5). Expression of inPAK, but not nonPAK, decreased MV-induced activation of PAK-1. (B) Representative fluorescence microscopy images of inPAK (iv, v, and vi)- or nonPAK (vii, viii, and ix)-transfected 3T3.CD4 cells at 24 h following infection with vMyxgfp (MOI of 1). Red and green cells indicate transfected and MV-infected cells, respectively, while yellow cells following merging of red and green fields indicate concurrent infection and transfection. Images obtained following merging of red and green fields from uninfected and nontransfected cultures (i), cultures only infected (ii), and cultures only transfected (iii) are included as controls. A prominent cell that is both infected and transfected with inPAK is indicated by an arrowhead. Original magnification, ×10. (C) Graphical representation of the number of cells concurrently infected and transfected with inPAK, nonPAK, or vector lacking exogenous sequences (mock). Values represent the mean number of yellow cells ± standard deviation (SD) from six fields in each of triplicate wells once the red and green fields were merged.