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. 2019 Jul 16;9:10278. doi: 10.1038/s41598-019-46531-y

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© The Author(s) 2019

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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PPP, fluvastatin or PD166866 do not inhibit 2D proliferation of JB6 P⁺ cells. Histograms of JB6 P⁺ cells stained with Cell Proliferation Dye eFluor™ 450 were treated with (A) PPP, (B) fluvastatin, PD166866 (C) or the vehicle, DMSO. Inhibition of proliferation is indicated by higher fluorescent signal that is represented by visually distinct rightward shifted histograms as shown following treatment with mitomycin C (MmC) and cycloheximide (CHX). The same histograms for DMSO, MmC (1.0 µM), and CHX (10 µg/mL) are shown for comparison with PPP, fluvastatin, and PD166866. (D) The eFluor™ 450 MFI of JB6 P⁺ cells (singlet, SYTOX Red negative cells) stained with Cell Proliferation eFluor™ 450 and treated with PPP, fluvastatin, or PD166866 was graphically depicted. PPP, fluvastatin, and PD166866 did not significantly influence the eFluor™ 450 MFI compared to vehicle (DMSO) controls, whereas MMC and CHX (positive controls) had significantly higher eFluor™ 450 MFI values. (E) PPP, fluvastatin, or PD166866 did not significantly decrease the % of live cells (singlet, SYTOX Red negative cells). MmC and CHX significantly induced cell death at 0.1 and 1.0 µM and 10 and 50 µg/mL respectively. Each treatment group had two replicates and analyzed by one-way ANOVA with multiple comparisons (**p < 0.01; ***p < 0.001).