Fig. 4

Host strain affects the moonlighting function (transferrin binding) of rGAPDH: far western blotting demonstrates that binding of recombinant GAPDH to transferrin varies with the host strain. a MT-GAPDH-H, b MT-H-GAPDH, c MS-GAPDH-H and d EC-H-GAPDH. The interaction of GAPDH with transferrin was detected using polyclonal rabbit α GAPDH antibody. rGAPDH from each source represents the positive control while BSA has been utilized as a negative control. Co-immunoprecipitation analysis comparing capture of: e MT-GAPDH-H and EC-H-GAPDH, f MT-GAPDH-H and MS-GAPDH-H. g MT-GAPDH-H and MT-H-GAPDH Blots were probed with polyclonal rabbit α GAPDH antibody (to determine capture) and α transferrin antibody (to confirm that equal amounts of transferrin were used). h Densitometry analysis, co-immunoprecipitation of MT-GAPDH-H was taken as 100 %. Experiments were repeated multiple times, data is represented as a histogram depicting % binding ± SD, n = 3, ***p ≤ 0.001