Characterization of C. elegans SAS-6 Protein Fragments and Depletion of Endogenous HsSAS-6 Using siHsSAS-6-3′UTR, Related to Figure 2 and Figure 3
(A) ITC of the C. elegans N-N interaction. Top panel: raw data representing the response to injections of ceN at high concentration into sample buffer. Bottom panel: integrated heat change (closed squares) and associated curve fit (black solid line).
(B) CD spectrum of ceN (open circles) or ceN[I154E] (closed circles) fragments.
(C) SDS-PAGE showing final purification products for AUC of ceN and ceN[I154E] recombinant proteins.
(D) SDS-PAGE showing final purification products for AUC of ceN-CC and ceN-CC[I154E] recombinant proteins.
(E) Cells left untreated (-), transfected with LO negative control siRNA (LO) or siHsSAS-6-3′UTR (si) for 48h before Western blot analysis with HsSAS-6 antibody; tubulin served as loading control. Arrows point to endogenous HsSAS-6 or HsSAS-6-GFP proteins. Dots indicate endogenous HsSAS-6 bands, stars unspecific bands.