Fig. 1
From: Molecular features of the UNC-45 chaperone critical for binding and folding muscle myosin
Producing functional myosin in insect cells. a Overview of constructs used for expression studies in insect cells. Western blot analysis of co-expression of NMY-2 (b) or MHC-B (c) with indicated chaperones in Hi5 insect cells. Whole cell lysates (WCL) and soluble fractions (SOL) are shown. 1/3rd of the WCL was applied with respect to the SOL fraction. (SBD–substrate binding domain, NBD–nucleotide binding domain, NTD–N-terminal domain, MD–middle domain, CTD–C-terminal domain). d Pull-down (PD) of Strep-tagged UNC-45 constructs from insect cells. Input samples and elutions were probed with Hsp70, Hsp90, and Strep-tag antibodies. e Upper panel: SEC traces of MHC-B purified from co-expression with the indicated C. elegans chaperones. Lower panel: SDS-PAGE gels showing the elution after NiNTA and indicated SEC fractions
