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. Author manuscript; available in PMC: 2009 Jan 11.
Published in final edited form as: Cell. 2008 Jan 11;132(1):137–148. doi: 10.1016/j.cell.2007.11.024

Figure 4. The Microtubule-binding Domain of SNPH Is Sufficient to Immobilize Axonal Mitochondria.

Figure 4

(A) SNPH associates with microtubules. COS cells were transfected with either GFPSNPH (1-469), a truncated SNPH in which the C-terminal mitochondrial targeting domain was deleted, or GFP-SNPH (1-469)ΔMTB, followed by staining for β-tubulin one day after transfection. In the middle pane, the transfected cells were treated with 10 μM of Nocadazol for 1 hr to disassemble microtubules.

(B) The microtubule-binding domain is required for SNPH to be co-pelleted with microtubules in a spin down assay. GST-SNPH (1-469) or GST-SNPH (1-469)ΔMTB was incubated in the absence or presence of Taxol-stabilized microtubules. The supernatant (S) and pellets (P) were analyzed by SDS-PAGE and visualized by Coomassie blue staining.

(C, D) Schematic diagrams and mobility of axonal mitochondria labeled with GFP-TOM22 or GFP-MTB-TOM22, a chimeric transgene in which the MTB was placed between GFP and the mitochondrial outer-membrane protein TOM22. Kymographs: 10 min. Scale bars: 10μm.