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. 2012 Apr 27;287(25):21406–21415. doi: 10.1074/jbc.M112.370213

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© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 5. — TIM family proteins are not part of the TGD complex. A, immunoblots of isolated chloroplast protein equivalent to 5 or 10 μg of chlorophyll were probed with At3g49560 antiserum (I) or its preimmune serum (P), as indicated at the top. B, antiserum raised against proteins encoded by At3g49560 was tested for specificity against the preprotein and amino acid transporter family. Full-length recombinant proteins for Tim17-1 (At1g20350), Tim17-2 (At2g37410), Tim17-3 (At5g11690), Tim23-1 (At1g17530), Tim23-2 (At1g72750), Tim23-3 (At3g04800), Tim22-1 (At1g18320/At3g10110), At3g25120, At3g49560, and At5g24650 (indicated at the top) were detected with antiserum recognizing proteins indicated at the right. C, chloroplasts isolated from wild-type Arabidopsis were separated by 4–14% HDN-PAGE in the first dimension and 12% SDS-PAGE in the second and then immunoblotted and detected with antisera recognizing proteins indicated at the right. D, immunoprecipitations of HA-tagged proteins, similar to those in Fig. 1E, were immunoblotted and probed with antisera recognizing proteins indicated at the right. C, chloroplast starting material; U, unbound fraction; W, final wash; E, eluate.