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Figure 6.
Figure 6. Localization of GFP-Tagged Skap-hom Variants in
Skap-hom^−/− BMMs
Skap-hom^−/− BMMs were
reconstituted with (A) GFP-tagged WT Skap-hom, (B) GFP alone, or
(C–G) the indicated GFP-tagged variants of Skap-hom. (H–N)
Cells were fixed and counterstained with rhodamine-labeled
phalloidin to visualize the actin cytoskeleton. (O–U) Merged
GFP and Rhodamine channels for each infection. WT Skap-hom (A,
H, and O) colocalizes with actin-rich ruffles. The DM^*
dimerization domain triple mutant (C, J, and Q) and the R140M PH
domain mutant (D, K, and R) are diffusely localized. Deletion of
the PH domain (ΔPH; [E], [L], and [S]) results in a protein
that localizes to actin ruffles. Skap-hom bearing both the R140M
and either the D129K (D129K:R140M, [F], [M], and [T]) or the
K56A (K56A:R140M, [G], [N], and [U]) mutations colocalize with
actin ruffles.
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