Fig. 2

SUMO-dependent interaction between FAF1 and SAP130. A Yeast strain L40 was co-transformed with bait (human WT SAP130 or mutant at three sumoylation residues (3KA), or the control protein lamin fused to the LexA DNA-binding domain) and prey constructs [human FAF1 WT, FAF1 with SIM mutant (DM), mSin3A or HDAC1 fused to the Gal-activation domain (AD)]. Yeast transformants were spotted on plates with histidine (-TULL) or without histidine (-TULLH), and with X-Gal (+ X-Gal) media. Schematic presentation of FAF1 DM and SAP130 C-terminal 3KA mutants analyzed in a yeast two-hybrid assay (top). B Yeast co-transformed with the indicated bait and prey were analyzed by quantitative β-Gal assays. C Coimmunoprecipitation assays. COS-1 cells co-transfected with Flag-FAF1 and HA-SAP130 WT or 3KA mutant of expression constructs were subjected to IP experiments followed by WB analysis with the indicated antibodies. WCL indicates whole cell lysate. D COS-1 cells co-transfected with myc-tagged mSin3A and HA-SAP130 WT or 3KA expression constructs were subjected to IP and WB analysis with the indicated antibodies. The immunoblots were cropped for clarity. Full length blots are presented in Supplemental Figure S9