FIGURE 8:

The GSK-3 inhibitor CHIR99021 increases the abundance of CREB-HΔTMC. (a) Cells transfected with the vector for CREB-HΔTMC under the control of the TK promoter were untreated or treated for 1 h (lanes 1–3) or 2 h (lanes 4 and 5) with 20 mM of NaCl or LiCl. (b) Cells were transfected with the vector for CREB-HΔTMC under the control of the CMV promoter and treated with either NaCl or LiCl for 1 h. Equal cell equivalents were analyzed for levels of CREB-HΔTMC using anti-SV5 or anti-pS73 antibodies. (c) Transfected cells were incubated in reduced serum (5% NBCS) 16 h before treatment with CHIR99021 (10 μM), LY294002 (10 μM), SB216763 (20 μM), TBCA (10 μM), or rapamycin (1 μM) for 24 h. Equal cell equivalents were analyzed for expression levels of CREB-HΔTMC using anti-SV5 antibody. (d) Transfected cells were incubated and treated as before with CHIR99021 (5, 10 μM) and LY294002 (5, 10 μM). Extracts were analyzed for total levels of CREB-HΔTMC using the anti-SV5 or for the pS73-phosphorylated form as before.