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. 2021 May 24;11:10814. doi: 10.1038/s41598-021-90184-9

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© The Author(s) 2021

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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Isolation and characterization of human PMCs from pleural lavage. (A) Representative photomicrograph of confluently growing cells obtained from pleural lavage in PSP patient. (B) Representative fluorescence-activated cell sorting dot plots of cultured cells. The left panel shows plots of the forward-scattered (horizontal) versus side-scattered (vertical) axes. The right panel represents the fluorescent intensities of APC (conjugated with the anti-podoplanin antibody) and FITC (conjugated with the biotinylated anti-mesothelin antibody), respectively. The Q2 area indicates the gateway for sorting PMCs (podoplanin-positive and mesothelin-positive cells). (C) The expression of MC markers (calretinin, WT1, keratin 5, and vimentin) examined in PSP-PMCs by RT-PCR. GAPDH was amplified as a control gene for RT-PCR. Gel electrophoresis of amplified products showed a discrete band with expected fragment size from each reaction. (D) SEM image of PSP-PMCs showing numerous microvilli characteristic of MCs. Scale bars represent 200 μm (A) and 20 μm (D), respectively. APC allophycocyanin, bp base pair, FITC fluorescein isothiocyanate, FSC-A forward scatter area, GAPDH glyceraldehyde 3-phosphate dehydrogenase, SSC-A side scatter area, WT1 Wilms’ tumor 1.