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. 2023 Oct 31;13:18736. doi: 10.1038/s41598-023-45470-z

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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PDGF-BB mediated SMC phenotypic switch induces interaction and nuclear expression of apelin and S100A4. (A) Double immunofluorescence staining showing apelin, S100A4, and both in S-SMCs after treatment with PDGF-BB for 96 h (n = 4). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (B) Graphs showing quantification of nuclear versus cytoplasmic apelin and S100A4 in S-SMCs treated with PDGF-BB vs control. Black and red lines were generated from a second order polynomial non-linear fit regression. (C) Data were separately displayed for a threshold value below or above 350 CCD units/5 s, and were analysed by simple linear regression and Pearson’s correlation. (D) Representative images of apelin and S100A4 interaction detected by in situ proximity ligation assay (PLA) in non-stimulated (Basal) and PDGF-BB stimulated SMCs (PDGF-BB). The control condition represents the absence of one of the primary antibodies (here the rabbit anti-apelin, Control). Scale bar, 20 μm. (E) Statistical analysis of average PLA complexes per cell (n = 4), Mann–Whitney non-parametric test, *p = 0.0286.