The hDOT1L interaction domain of CALM-AF10 is required for the induction of H3K79 hypomethylation. (A) Coimmunoprecipitation of CALM-AF10 with hDOT1L depends on the OM-LZ domain of AF10. HA-tagged hDOT1L was coexpressed with Flag-tagged CALM-AF10 or its deletion mutant, as indicated in 293T cells, and the cell lysate was subjected to coimmunoprecipitation using Flag antibody. The presence of hDOT1L-HA in the immunoprecipitate was examined by Western blotting. One percent of the input cell lysate was loaded (In, input; FT, flow through; IP, immunoprecipitation). The expression of the AF10 fusion proteins in the input lysate was confirmed with Western blotting using a Flag antibody (bottom panel). (B) Expression of CALM-AF10 fusion protein lacking the OM-LZ domain failed to reduce H3K79 methylation in T-REx-293 cells. The tetracycline-induced expression of Flag-tagged fusion proteins was confirmed (top panel).