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. 2019 Feb 26;294(16):6494–6505. doi: 10.1074/jbc.RA118.006071

Figure 5.

Figure 5.

The SH3 domain determines recruitment of IRTKS to RAB11. A, schematic of MIM, IRTKS, and their fusion mutants. WH2, WASP homology 2. B, HeLa cells expressing GFP-tagged IRTKS or IRTKSΔSH3 proteins were stimulated with 150 ng/ml SDF-1 for 30 min. The cell lysates were subjected to immunoprecipitation (IP) with GFP antibody, followed by immunoblot (IB) using anti-RAB11, RAB5, or RAB7, as indicated. C, cells expressing GFP-tagged MIM, IRTKS, and IRTKSΔSH3 were treated with 500 ng/ml SDF-1 for up to 45 min. The levels of surface CXCR4 were estimated by flow cytometry. The data represent mean ± S.E.M. (n = 3). *, p < 0.05; ***, p < 0.001; referring to the differences between cells expressing IRTKS-GFP and those expressing IRTKSΔSH3-GFP. D, cells expressing MIM-GFP or its fusions were treated with 150 ng/ml SDF-1 for 30 min. The interactions of MIM and its fusions with RAB7 and RAB11 were analyzed as described in B.