Figure 6. Immunoblot analyses to assess interaction of YidC with respiratory proteins.

(a,b) M. smegmatis lysate expressing YidC of Mtb was incubated individually with QcrA-FLAG, cMyc-Rv0247c and Rv0248c-FLAG-expressing lysates for 16 hours followed by immunoprecipitation (IP) using anti-FLAG, anti-cMyc or anti-YidC antibodies, respectively. The immunoprecipitates of anti-FLAG or anti-cMyc antibodies were subjected to immunoblotting (IB) by anti-YidC (a), whereas those immunoprecipitated by anti-YidC were probed by anti-FLAG or anti-cMyc antibodies (b), respectively. Blots were developed by using clean-blot IP detection kit (Thermo Pierce) to avoid signals from denatured IgG subunits present in the immunoprecipitated samples. Data represent two experiments in (a,b).