Skip to main content
. 2014 Sep 19;9(9):e107914. doi: 10.1371/journal.pone.0107914

Figure 1. USF2 is phosphorylated by GSK3β.

Figure 1

(A) USF2 was immunoprecipitated from GSK3β+/+ and GSK3β−/− cells and phospho-USF2 protein levels were detected with phospho-threonine or phospho-serine antibodies. (B) USF2 was immunoprecipitated from HeLa cells transfected with pcDNA3-GSK3β-WT-HA. Following SDS-PAGE phosphoproteins were visualized with the Pro-Q Diamond Phosphoprotein Gel Stain. The total protein amount was detected by silver staining and GSK3β expression was verified by Western blotting. (C) Where indicated, HeLa cells transfected as above were treated with the GSK3 selective inhibitors BIO (1 µM), LiCl (10 mM), 1-Azakenpaullone (Aza, 7.5 µM) for 1 h. Proteins were isolated 24 h after transfection and detected by Western blotting. (D) Cells were transfected with expression vectors for USF2 and GSK3β and the cell extract was incubated with calf intestinal phosphatase (CIP) or only with buffer (Mock). Proteins were detected by Western blotting.