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Fig. 5. Dynamin and eNOS are assembled into common structures by NOSTRIN in a process requiring its SH3 domain. CHO-eNOS cells stably expressing eNOS were transiently transfected with cDNA encoding NOSTRIN (A-D), dynamin-2-GFP (E-H), NOSTRIN and dynamin-2-GFP (I-L), or NOSTRIN-{Delta}SH3 and dynamin-2-GFP (M-P). Following fixation, cells were stained with anti-NOSTRIN (B,J,N) and anti-eNOS (C,G,K,O). (E,I,M) Fluorescence of GFP-fused dynamin-2. (D,H,L,P) Merged images of each row. (A-D) NOSTRIN and eNOS co-localize at vesicular and elongated structures throughout the cytoplasm. (E-H) In the absence of NOSTRIN, eNOS and dynamin-2-GFP hardly overlap. (I-L) Upon co-expression of dynamin-2-GFP and NOSTRIN, dynamin and eNOS are brought together at common structures. (M-P) NOSTRIN-{Delta}SH3 does not affect localization of either eNOS or dynamin-2-GFP. Bars, 10 µm.





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