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. 2022 Sep 28;45(6):1311–1328. doi: 10.1007/s13402-022-00719-z

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© The Author(s) 2022

Open AccessThis 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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Fig. 1 — Effect of WPMY-1 prostate fibroblasts on PC3 cell proliferation and migration and application of CTAP to this model system.A and B, WPMY-1 co-culture increases the proliferation and migration of PC3 cells. Upper panels (A) – images of GFP-labelled PC3 cells after mono- or co-culture. The graph provides image quantification. Results of random cell migration assays for PC3 mono-culture and WPMY-1 co-culture (B). C and D, WPMY-1 conditioned medium increases the proliferation and migration of PC3 cells. Results of MTS assay for PC3 cells with either basal medium, full medium or WPMY-1-conditioned medium (C) and Transwell assays for PC3 cells with either basal medium or WPMY-1-conditioned medium (D). Data are presented as mean ± SD of 3 biological replicates; student t-test, **p < 0.01, ***p < 0.001, ****p < 0.0001. Scale bar: 1 mm. E, Application of CTAP to the PC3/WPMY-1 model. CTAP provides spatial resolution, identifying which cell type the MS-detected peptides are derived from. Discrimination between additional variables is achieved by differential isobaric tandem mass tag (TMT) labelling of peptides. F, Generation of PC3 and WPMY-1 cells expressing HA-tagged Lyr and DDC, respectively. Cell lysates were Western blotted as indicated. CM, conditioned medium