The ratio of SPFH1 to SPFH2 in the SPFH1/2 complex is flexible.
A, lysates from Rat-1 cells expressing the indicated shRNAs were
prepared in 1% Triton X-100-containing lysis buffer, were subjected to
BN-PAGE, and were probed for SPFH1 or SPFH2. B, anti-SPFH1 or
anti-SPFH2 IPs from Rat-1 cells expressing the indicated shRNAs were Coomassie
Blue stained, and bands corresponding to SPFH1 and SPFH2 are marked.
C, HeLa cells were transfected with varying amounts of SPFH1-FLAG and
SPFH2-HA cDNA, anti-HA IPs were Coomassie Blue stained, and bands
corresponding to SPFH1-FLAG and SPFH2-HA are marked. IP,
immunoprecipitate; IB, immunoblot.