Figure 1.

Validation of specificity of anti-RNF11 polyclonal antibody. A, validation of affinity-purified RNF11 rabbit polyclonal antibody by knockdown in 293T cells. Shown is immunoblotting of 20 μg of solubilized P20K pellet. Inputs were resolved by SDS-PAGE and blotted. The blot was then cut in half at the 37 kDa marker. The top and bottom halves of the blot were probed with tubulin antibody and then developed together. Inputs show a decrease in RNF11 signal in cells stably expressing an shRNA specific for RNF11. α-Tubulin was used as a loading control. B, endogenous and C-terminally tagged RNF11 localize to the endosomal compartment. HeLa cells were transfected with a C-terminal FLAG fusion with RNF11 mammalian expression vector, fixed after 48 h, and visualized with FLAG antibody and the early endosomal marker EEA1 (bottom). Endogenous RNF11 immunostaining with the in-house RNF11 antibody and EEA1 antibody is shown in the top panel.