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. 2002 Oct;22(20):6946–6958. doi: 10.1128/MCB.22.20.6946-6958.2002

FIG. 5.

FIG. 5.

Mislocation of Rsp5p in mutants with defective actin cytoskeletons. (A) Rsp5p is localized in clumps that contain F-actin in the arp2-1 mutant. The arp2-1 strain was transformed with YCpHA-RSP5 or YCpHA-rsp5-w2, and the transformants were processed for immunofluorescence using anti-HA antibody and stained with phalloidin. The structures containing HA-Rsp5p and actin are indicated by arrows. (B) Sla2p and Rsp5p colocalize in arp2-1 mutant cells. Transformants were double stained with anti-HA and anti-Sla2p antibody. The structures containing HA-Rsp5p and Sla2p are indicated by arrows. (C) Rsp5p is present in clumps that contain actin in pan1-9 cells. Plasmids bearing HA-RSP5 or HA-rsp5-w were transformed into the pan1-9 mutant. Cells were stained by indirect immunofluorescence using the anti-HA antibody and subsequently stained with phalloidin. The clumps of concentrated HA-Rsp5p colocalizing with actin are indicated by arrows. Some normal actin patches that colocalize with punctate structures of HA-Rsp5p are indicated by arrowheads. Actin clumps are present in almost every cell expressing HA-rsp5-w2 (bottom panel). (D) Sla2p is present in the same clumps as Rsp5p in the pan1-9 mutant. Cells expressing HA-rsp5-w2 were stained by double indirect immunofluorescence using anti-HA and anti-Sla2p antibody. The dots that correspond to HA-Rsp5-w2p and Sla2p are indicated by arrows.