Fig. 2. Measurement of the interaction of Mlph with EB1 and MyoVa-MSGTA in vitro. Panels A and B show interaction with EB1 and MSGTA involves different regions of Mlph (A) and that Mlph may interact simultaneously with EB1 and MSGTA (B). One hundred pmol of GST-Mlph protein (A) or MBP-EB1 (B) was incubated with equimolar amounts of MBP-EB1 or MBP-MSGTA (A) or his₆-Mlph and/or his₆-MSGTA (B) in the presence of glutathione-sepharose (A) or amylose agarose (B) as described in the Materials and Methods. Bound proteins were precipitated, eluted from the beads, and subjected to immunoblotting using indicated antibodies. In A, relative binding (see bar chart) was calculated by dividing band intensity of EB1 or MSGTA by the band intensity of Mlph (as described in Materials and Methods). Experiments were carried out in triplicate and the migration of molecular-weight standards is indicated on the left of each blot.