Figure 4. Co-immunoprecipitation analysis on endogenous expression level.

HEK293T cell lysates were incubated with Protein G beads loaded with either anti-AMOT or IgG isotype control antibody. After 2 h of incubation, the beads were washed twice in lysis buffer and protein complexes were eluted from the beads in SDS-PAGE loading buffer. Immunoblots were sequentially incubated with anti-AMOT and anti-NEMO antibody. Immunoprecipitation (IP) of AMOT (both p80 and p130 isoforms) and co-immunoprecipitation of NEMO is clearly visible. One experiment of three biological replicate experiments is shown. Full Western blots of all three replicate experiment and antibody validation can be found in the Figshare repository (Uncropped Western blots, Data Citation 2).