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. 2013 Aug 5;110(34):E3162–E3170. doi: 10.1073/pnas.1312531110

Fig. 5.

Fig. 5.

Interaction of in vitro-synthesized BIG2 with BIG1 and NMHC IIA, but not MYPT1 or PP1cδ. BIG2, full-length (FL) and three fragments (BIG2-N, -S, and -C) with N-terminal HA tags and other proteins were synthesized singly in wheat germ extracts and mixed before IP and analyses of collected proteins. (A) After mixing BIG2 and NMHC IIA, samples before (In, 5%) and from IP with control IgG or anti-BIG2 antibodies (50%) were reacted with antibodies against BIG2 or NMHC IIA. (B) HA-tagged BIG2 fragments were mixed with NMHC IIA before IP with antibodies against NMHC IIA for analysis of collected proteins. Arrow indicates BIG2-N, arrowhead BIG2-S, and asterisk BIG2-C. (C and D) BIG2, MYPT1, and PP1cδ were mixed as indicated before IP with control IgG or antibodies against BIG2 that failed to collect MYPT1 (C) or PP1cδ (D). (E) BIG2, c-myc-MYPT1, and HA-BIG1 were mixed before IP with BIG2 antibodies that collected both MYPT1 and HA-BIG1. (F) IP of BIG2 after incubation of BIG2, PP1cδ, and BIG1 yielded PP1cδ with BIG2 and BIG1.