FIGURE 7:

Inhibition of GSK-3 with CHIR99021 stimulates ER–Golgi trafficking of CREB-H. (a, b) Subcellular localization of CREB-H in normal S1P/S2P background (a) or S1P/S2P-ve backgrounds (b). COS-1 cells were transfected with a plasmid expressing wt CREB-H or the S1P⁻/S2P⁻ form under the control of a TK promoter. Medium was replaced at 32 h with medium containing 5% serum for 16 h, followed by treatment without (I and III) or with CHIR99012 (II and IV) added at 10 μM for 6 h. Cells were fixed in 4% paraformaldehyde and stained with anti-SV5 (red channel) and anti-TGN46 (green channel) primary antibodies. Arrows indicate features as discussed in the text.