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Fig. 4 | Cell Communication and Signaling

Fig. 4

From: SUMO3 modification by PIAS1 modulates androgen receptor cellular distribution and stability

Fig. 4

PIAS1 itself was modified by SUMO3 at 117th lysine residue. (a) DU145 cells were cotransfected with plasmids as indicated for 48 h. Whole-cell lysates were immunoprepapited with anti-AR antibody and then analyzed by immunoblot analysis using the indicated antibodies against AR (IP, top panel) and PIAS1 (IB, second panel). Whole-cell lysates (Input) were also immunoblotted with anti-AR (third panel), anti-PIAS1 (fourth panel) or anti-actin (bottom panel) antibodies. (b) DU145 cells were co-transfected with plasmids as indicated for different transfection periods (24 h, 48 h, 72 h and 96 h). Whole cell lysates generated together were immunoprepapited with anti-myc antibody. The myc-immunoprepapite was then detected by anti-myc (IP, top panel) and anti-SUMO3 immunoblotting (IB, second panel). Whole-cell lysates (Input) were also immunoblotted with anti-myc (third panel) or anti-actin (bottom panel) antibodies. (c) Diagrammatic representation of putative typical sumoylation site on human PIAS1 sequense predicted by using the GPS-SUMO software. Analysis of human PIAS1 sequence indicated the presence of only one putative typical sumoylation site, Lys-117, which located near to the PINIT domain of PIAS1. (d) DU145 cells were co-transfected with plasmids as indicated for 48 h and Whole cell lysates were immunoprepapited with anti-myc antibody. The myc-immunoprepapite was then detected by anti-myc (IP, top panel) and anti-SUMO3 immunoblotting (IB, second panel)

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