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. 2007 May;18(5):1609–1620. doi: 10.1091/mbc.E06-06-0518

Figure 3.

Figure 3.

ScarΔ96-GFP is stably expressed and complexes differently than wild-type Scar. (A) Growing HPS400 cells expressing Scar (∼66 kDa) and/or ScarΔ96-GFP (∼75 kDa) were collected and Western blotted for Scar. ScarΔ96-GFP is stably expressed in both backgrounds. Control cells are HPS400 and Scar null cells expressing empty vector. Clarified cell lysate from parental cells expressing ScarΔ96-GFP was loaded onto a 7–20% sucrose gradient. Fractions of 0.5 ml, were collected and immunoblotted for Scar (B) or PIR121 (C). Numbers above blots indicate fraction number. The majority of endogenous Scar is present in fractions 9–12 as in Figure 1. ScarΔ96-GFP is present in fractions 14–18 (B), which contain protein complexes of ∼150–300 kDa. PIR121 protein is not detected in the fractions containing ScarΔ96-GFP but continues to track with wild-type Scar (C). (D) Cells null for each of the Scar complex members with or without expressed ScarΔ96-GFP were collected and Western blotted as in A. ScarΔ96-GFP is stably expressed in each background.