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. 2002 Nov;22(22):7993–8004. doi: 10.1128/MCB.22.22.7993-8004.2002

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Copyright © 2002, American Society for Microbiology

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FIG. 9. — Association of Pf20 with Spag6. (A) Spag6-RFP induces Pf20-GFP localization to microtubules in cotransfected CHO cells. Living CHO cells were cotransfected with Spag6-RFP and Pf20-GFP. In cells that were coexpressing both Spag6-RFP and Pf20-GFP, Spag6-RFP (subpanel B) was found to be exclusively colocalized with Pf20-GFP (subpanel A) in a subset of microtubules. The overlap is shown in subpanel C. In living CHO cells that were transfected with Pf20-GFP alone (subpanel D), Pf20-GFP was found dispersed in a cytoplasmic pattern and no Pf20-GFP labeled microtubules were observed in the transfected cells. All are confocal images. Images A, B, and C are projections of eight z-section series, obtained with simultaneous dual-color scanning (bars = 10 μm). (B) A portion of Pf20 WD-GFP binds to Spag6-RFP-labeled microtubules in cotransfected CHO cells. (Subpanels A to C) Living CHO cells cotransfected with Spag6-RFP and Pf20 WD-GFP. In cells coexpressing both constructs, Pf20 WD-GFP (subpanel A) was found to colocalize to Spag6-RFP-labeled microtubules (subpanel B). Arrowheads in insets in subpanels A, B, and C show colocalization of Pf20 WD-GFP (subpanel A) and Spag6-RFP (subpanel B) in microtubules. Subpanel C is a merged image of subpanels A and B. Pf20 WD-GFP (subpanel D) is present in a cytoplasmic and nuclear pattern when expressed alone in CHO cells, and no Pf20 WD-GFP-labeled microtubules were found in the transfected cells. All are confocal images. Images A, B, and C are projections of eight z-section series, obtained with simultaneous dual-color scanning (bars = 10 μm). (C) Western blot analysis of Pf20 in extracts of transfected COS-1 cells. COS-1 cells were transfected with mouse Pf20 cDNA (pEGFP/Pf20) or empty vector. (Subpanel A) Total extract of COS-1 cells showing expression of Pf20-GFP in cells transfected with Pf20-GFP cDNA but no expression in nontransfected COS-1 cells or cells transfected with the empty plasmid vector. (Subpanel B) The same as subpanel A, except that the antibody was blocked by the purified protein. Subpanel C shows that COS-1 cells transfected with pEGFP vector and pEGFP/Pf20 express the GFP.

FIG. 9. — Association of Pf20 with Spag6. (A) Spag6-RFP induces Pf20-GFP localization to microtubules in cotransfected CHO cells. Living CHO cells were cotransfected with Spag6-RFP and Pf20-GFP. In cells that were coexpressing both Spag6-RFP and Pf20-GFP, Spag6-RFP (subpanel B) was found to be exclusively colocalized with Pf20-GFP (subpanel A) in a subset of microtubules. The overlap is shown in subpanel C. In living CHO cells that were transfected with Pf20-GFP alone (subpanel D), Pf20-GFP was found dispersed in a cytoplasmic pattern and no Pf20-GFP labeled microtubules were observed in the transfected cells. All are confocal images. Images A, B, and C are projections of eight z-section series, obtained with simultaneous dual-color scanning (bars = 10 μm). (B) A portion of Pf20 WD-GFP binds to Spag6-RFP-labeled microtubules in cotransfected CHO cells. (Subpanels A to C) Living CHO cells cotransfected with Spag6-RFP and Pf20 WD-GFP. In cells coexpressing both constructs, Pf20 WD-GFP (subpanel A) was found to colocalize to Spag6-RFP-labeled microtubules (subpanel B). Arrowheads in insets in subpanels A, B, and C show colocalization of Pf20 WD-GFP (subpanel A) and Spag6-RFP (subpanel B) in microtubules. Subpanel C is a merged image of subpanels A and B. Pf20 WD-GFP (subpanel D) is present in a cytoplasmic and nuclear pattern when expressed alone in CHO cells, and no Pf20 WD-GFP-labeled microtubules were found in the transfected cells. All are confocal images. Images A, B, and C are projections of eight z-section series, obtained with simultaneous dual-color scanning (bars = 10 μm). (C) Western blot analysis of Pf20 in extracts of transfected COS-1 cells. COS-1 cells were transfected with mouse Pf20 cDNA (pEGFP/Pf20) or empty vector. (Subpanel A) Total extract of COS-1 cells showing expression of Pf20-GFP in cells transfected with Pf20-GFP cDNA but no expression in nontransfected COS-1 cells or cells transfected with the empty plasmid vector. (Subpanel B) The same as subpanel A, except that the antibody was blocked by the purified protein. Subpanel C shows that COS-1 cells transfected with pEGFP vector and pEGFP/Pf20 express the GFP.