Figure EV3. AMOTL1 promotes Merlin ubiquitination.

• A
  Merlin‐depleted LN229 cells stably expressing Flag‐HA-tagged Merlin used in Fig 4D were stably transduced with indicated shRNAs targeting AMOTL1 or a scrambled shRNA and subjected to Western blotting.
• B
  HEK293T cells were transfect with indicated genes and subjected to nickel pull‐down in a native condition followed by Western blotting. Myc‐Merlin (arrow); non‐specific bands (asterisk).
• C
  Merlin‐depleted LN229 cells stably transduced with Flag‐tagged wild‐type Merlin or the indicated mutants were subjected to immunoprecipitation with a Flag antibody. The lysate and immunoprecipitated products were subjected to Western blotting.
• D, E
  HEK293T cells were transfected with indicated genes and subjected to Western blotting. The ratio of mono‐ubiquitinated to native Merlin in each lane was quantified by ImageJ and is shown under the blot in (D).

Source data are available online for this figure.