FIGURE 6.

Both TgMLC1 and TgELC1 are bound to the same heavy chain. TgMyoA heavy chain (HC) and TgUNC were co-expressed in Sf9 cells with both 3xMyc-MLC1 and 3xHA-ELC1. The lane labeled S is the total soluble protein after Sf9 cell lysis. TgMyoA was co-immunoprecipitated from cell lysates using either anti-HA antibody (to immunoprecipitate TgELC1; left panels) or anti-TgMLC1 antibody (right panels), confirming that each of the light chains binds to TgMyoA heavy chain. Furthermore, TgMLC1 is present in the TgELC1 pulldowns and vice versa, demonstrating that the two light chains are simultaneously present on the same MyoA heavy chain. TgUNC (∼126 kDa) was detected with anti-Myc antibody, TgMyoA heavy chain (∼104 kDa) was detected with anti-FLAG, 3xMyc-TgMLC1 (∼29 kDa) was detected with anti-Myc in the left panels and anti-TgMLC1 in the right panels, and 3xHA-ELC1 (∼18 kDa) was detected with anti-HA. WB, Western blot; IP, immunoprecipitate.