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. 2022 Aug 23;54(8):1201–1213. doi: 10.1038/s12276-022-00815-y

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© The Author(s) 2022

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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Fig. 5 — a The top 15 MetaCore pathways (based on their minimum FDR) enriched for both the upregulated genes in Ang II (100 nM, 24 h)-stimulated rat aortic SMCs and the downregulated genes in PDE4D siRNA (200 nM, 48 h)-treated SMCs. b Representative immunoblot analysis of caspase-3 and cleaved caspase-3 expression in the SMCs treated with Ang II (100 nM, 24 h) and/or PDE4D siRNA (200 nM, 48 h) as indicated. c Quantification of cleaved caspase-3 protein expression by immunoblotting in (b) normalized to caspase-3 protein (fold change versus control). **p < 0.01, ***p < 0.001, two-way ANOVA with Holm-Sidak’s post hoc test, mean ± SEM, n = 3 separate experiments. d Flow cytometric analysis of annexin V/propidium iodide (PI)-stained SMCs treated with or without Ang II (100 nM, 24 h)/PDE4D siRNA (200 nM, 48 h) as indicated. e Quantification of the total (early and late) apoptosis rates of annexin V/PI-stained SMCs treated with or without Ang II (100 nM, 24 h)/PDE4D siRNA (200 nM, 48 h) as indicated. ****p < 0.0001, two-way ANOVA with Holm–Sidak’s post hoc test, mean ± SEM, n = 3 separate experiments. f Immunoblot analysis of caspase-3 and cleaved caspase-3 expression in the SMCs treated with or without Ang II (100 nM, 24 h)/rolipram (500 nM, 24.5 h). g Quantification of cleaved caspase-3 protein expression by immunoblotting in (f) normalized to caspase-3 protein (fold change versus control). **p < 0.01, two-way ANOVA with Holm–Sidak’s post hoc test, mean ± SEM, n = 3 separate experiments. h Flow cytometric analysis of annexin V/PI-stained SMCs treated with or without Ang II (100 nM, 24 h)/rolipram (500 nM, 24.5 h) as indicated. i Quantification of the total (early and late) apoptosis rates of annexin V/PI-stained SMCs treated with or without Ang II (100 nM, 24 h)/rolipram (500 nM, 24.5 h) as indicated. ****p < 0.0001, two-way ANOVA with Holm-Sidak’s post hoc test, mean ± SEM, n = 3 separate experiments.