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. 2017 Jun 1;28(11):1565–1579. doi: 10.1091/mbc.E17-01-0075

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© 2017 Barbosa et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 5: — GSK-3 inhibition induces cleavage of CREB-H and reduces phosphorylation at serines 73 and 81. (a) COS-1 cells transfected with the vector expressing the precursor form of CREB-H under the control of a TK promoter were untreated or treated with 20 mM of either NaCl or LiCl for 1 h (lanes 1–3), 2 h (lanes 4–6), or 4 h (lanes 7 and 8). Extracts of equal cell equivalents were analyzed by Western blot using the anti-SV5 antibody and anti–γ-tubulin as a loading control. (b, c) Extracts were analyzed by Western blot for the presence of phosphate at serines 73 and 81 of CREB-H using the anti-SV5, anti-pS73 (b, lanes 1–7), and anti-pS81 (c, lanes 1–7) antibodies as described in Materials and Methods. FL refers to the bands of the precursor form of CREB-H and Cl to the cleavage product of CREB-H. Asterisk indicates a nonspecific band detected by the antibody.