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. 2022 Jul 1;41(30):3804–3820. doi: 10.1038/s41388-022-02389-5

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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. 4 — A Western blot analysis of the indicated signaling proteins in PC3, PC3-PLEC-KO, PC3-α6-KO, PC3-PLEC-α6-dKO, PC3-β4-KO, and PC3-PLEC-β4-dKO cell lines. B Cell proliferation and C wound-closure analysis of PC3, PC3-PLEC-KO, PC3-PLEC-α6-dKO, and PC3-PLEC-β4-dKO cells using IncuCyte S3. The data shown is a representative experiment of three independent repeats showing the mean ± SD from a minimum of 4 replicates per sample. D Wound-closure analysis was performed for RWPE1, RWPE1-PLEC-KO, RWPE1-PTEN-α6-PLEC-tKO and RWPE1-PTEN-β4-PLEC-tKO cells as described in C. E Western blot analysis of apoptosis markers cleaved caspase-3 and cleaved PARP in PC3, PC3-α6-KO, PC3-PLEC-α6-dKO, PC3-β4-KO and PC3-PLEC-β4-dKO. F The indicated PC3 cell variants grown for 2, 4 or 7 days on polyHEMA-coated plates were harvested and subjected to analysis of anoikis resistance using the Annexin V/propidium iodide. The data shows the mean ± SD from three independent experiments. The different PC3 cell variants were grown for 21 days in soft agar followed by the determination of G the total area of colonies and H the average size of individual colonies. The data shows the mean ± SD from three independent experiments. I PC3, PC3-PLEC, PC3-α6-dKO, PC3-PLEC-α6-dKO, PC3-β4-KO, and PC3-PLEC-β4-dKO cells were grown for 72 h in the presence of the indicated concentrations of docetaxel followed by the analysis of cell viability using an MTT-assay. The data shows the mean ± SD from three independent experiments. J Comparison of the most enriched pathways in the indicated PC3 cell variants revealed by the RNA-Seq analysis. K The list of top-enriched pathways upregulated in PC3-α6-KO cells when compared with the parental PC3 cells. L Enrichment plots of the 1^st- and 5^th- ranked pathways in PC3-α6-KO cells. M Quantitative PCR analysis in the different PC3 cell variants to validate upregulation of selected plectin-independent and N plectin-dependent target genes of the G2M checkpoint pathway. The analysis was performed in triplicate and is presented as mean ± SD. All the western blot data in this figure are representative of at least three independent experiments. Asterisks indicate significance (p-value: *<0.05; **<0.01; ***<0.001).