Fig. 3.
Size-exclusion FPLC analysis of nuclear RA-binding activity in COS-1 cells.
Cells are transfected with pSG5 vector (A), wild-type PML-RARα L form (B), mutant Lys207Asn (C), mutant Arg272Gln (D), mutant Gly289Arg (E), mutant Pro407Ser (F), wild-type PML-RARα S form (G), and mutant Arg294Trp (H). Nuclear extracts were incubated with 10 nmol/L [3H]-RA in the absence (●) or presence (○) of 200-fold excess of unlabeled RA for 15 hours at 4°C. The samples were fractionated over a Superose 6HR 10/30 column at 0.4-mL intervals. Arrows indicate the fraction numbers of marker proteins (in kd) used to calibrate the FPLC column.