Additional file 7.
Negative controls of immunoprecipitations shown in Figure 4. Additional file 7 is a figure showing the test for unspecific binding of non-consensus C-terminally truncated proteins (MYC:MIRdCter) to irrelevant HA epitoped peptide in co-immunoprecipitation reaction corresponding to negative control of the experiment described in Figure 4. Each MYC:MIRdCter construct was co-expressed in 293T cells together with empty pDEST-CMV-3xHA vector and co-IPed using anti-HA sepharose beads. Binding of given protein upon precipitation was revealed by western blotting using anti-MYC serum. For each IP performed three panels are presented. Upper panel: IP reaction probed after resolution on SDS-PAGE and blotting with anti-HA antibody. Middle panel: the same IP reaction probed with anti-MYC serum detecting the truncated protein fragments (MYC:MIRdCter). Lower panel: detection of expression of each truncated protein fragment by probing total crude cellular extracts (input) with anti-MYC serum.
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Lenfant et al. BMC Genomics 2010 11:671 doi:10.1186/1471-2164-11-671