Western blot analysis of RasD protein level. Cell lysates from the JH10, rasG-, rasG-/[rasG]:rasD, rasC-/rasG-, rasC-/rasG-/[rasG]:rasD, rasC-, and rasD- strains were probed with RasD specific antibody. For the rasG-/[rasG]:rasD and the rasC-/rasG-/[rasG]:rasD cells, lanes “a,” “b,” and “c” refer to independently clonal isolates.