FIG. 5.

Release of LRH-1 from PML-NBs correlates with the loss of SUMO moiety. (A) HeLa cells were transfected with YFP-LRH-1, HA-SUMO, PIASxα, and PML-III expression vectors. (C) HeLa cells were transfected with YFP-SUMO-1, myc-LRH-1, PIASxα, and PML-III expression vectors. The cells were incubated in a 0.5% Triton X-100-containing buffer for 1 min before fixation or extraction. Triton X-100 treatments were performed in the presence or absence of 10 mM NEM and 10 μg of E64 isopeptidase inhibitors/ml. Confocal microscopic images (A and C) and Western blot analysis of extracts with αLRH-1 antibody (B) are shown.