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. Author manuscript; available in PMC: 2013 Dec 1.
Published in final edited form as: J Mol Endocrinol. 2013 Apr 23;50(3):361–374. doi: 10.1530/JME-12-0154

Figure 5.

Figure 5

Figure 5

Egr-1 synthesis is dependent on ERK1/2 transcriptional activation and de novo synthesis. (A) Western blot analysis of Egr-1 after 1 h treatment with 25 μM 13cRA, with or without the MEK inhibitor (20 μM) PD98059. (B) Western blot analysis of Egr-1 after 1 h treatment with 25 μM 13cRA, with or without the translational inhibitor cycloheximide (CHx). (C) Western blot analysis of nuclear extracts treated with and without 13cRA, probed with anti-Sp1 primary antibody after immunoprecipitation with anti-Egr-1. (D) Western blot analysis of nuclear extracts treated with and without 13cRA, probed with anti-Egr-1 primary antibody after immunoprecipitation with anti-Sp1. (E & F) Corresponding reprobe controls with appropriate capture antibody to confirm efficient immunoprecipitation of target proteins.