(A) Quantitation of the degree of overlap between the GFP-SNX5 constructs and the mCherry-Rab25 inclusion membrane marker from Figure 6A. Mander’s correlation coefficient of mChRab25 signal over GFP-SNX5 signals (10 cells per group; error bars, S.D). (B) Co-immunoprecipitation of GFP-SNX5 from HeLa cells shows that both the wild-type and mutant protein (F136A) interact equally with endogenous SNX1. This indicates that both proteins are correctly folded and otherwise functional. (C) HeLa cells stably expressing mCherry-Rab25 (red) were transfected transiently with GFP-IncE(91-132) or GFP-IncE(91-132)(F116D) (green) and infected with C. trachomatis L2 for 18–24 hr. Mutation in the SNX5 binding IncE peptide (F116D) abolishes recruitment to endosomal structures. Neither construct is recruited to inclusions, which is consistent with the lack of transmembrane regions. *Represents the inclusion. Scale bar 20 μm.