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. 2018 Mar 15;8:4655. doi: 10.1038/s41598-018-23092-0

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Functional role of MAPK cascades and PI3K/Akt/P70S6K pathway in PACAP-induced PRL promoter activity. αT3-1 cells were transiently transfected with pPRL(−1156).LUC for 6 h by using lipofectamine. After 18 h recovery, the cells were incubated with respective drugs. In this study, αT3-1 cells over-expressed pPRL(−1156).LUC were treated with oPACAP₃₈ (10 nM, 24 hrs) in the presence or absence of (A) ERK1/2 inhibitor U0126 (10 μM), (B) p38MAPK inhibitor PD169316 (10 μM), (C) JNK inhibitor SP600125 (10 μM), (D) PI3K inhibitor Wortmannin (10 nM), (E) P₇₀^S6K inhibitor rapamycin (20 nM) and (F) Akt inhibitor API-2 (10 uM). After drug treatment, cell lysate was prepared for dual-luciferase measurement. Data presented were expressed as percentage of control by conversing the ratio of firefly and renilla luciferase in the same sample. Data presented are expressed as mean ± SEM (n = 4) and different letters denote a significant difference at p < 0.05 (ANOVA followed by Fisher’s LSD Test).