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. 2017 Jan 23;13(1):e1006147. doi: 10.1371/journal.ppat.1006147

Fig 3. Complex formation occurs with the constitutively active SmVKR1 variant but not with a dead-kinase mutant.

Fig 3

Co-Immunoprecipitation of HA-tagged Smβ-Int1 (β-Int-HA) and a selection (see Fig 2; the same numbering was used, except lanes 14, 15) of combinations of V5-tagged SmILK (ILK-V5), SmPINCH (PINCH-V5), and SmVKR1 (VKR1-V5) constructs in Xenopus oocytes. Anti-HA antibodies immunoprecipitated Smβ-Int1 together with SmILK, SmPINCH, endogenous Nck2, and SmVKR1 upon co-expression in oocytes only when their wildtype forms (lane 11) or the constitutively active form of SmVKR1XE (lane 14) were used. In these cases Xenopus Nck2 was part of the complex formation as detected in lanes 11 and 14 by anti-human Nck2 antibody. Using deletion variants (DEL) of SmILK (lane 12) and SmPINCH (lane 13), SmILK individually (lane 1), a combination of wildtype SmILK, SmPINCH and Smβ-Int1 (lane 7), or a dead kinase variant (KO) of SmVKR1 (lane 15), no immunoprecipitated interaction-partners were detected.