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. 2011 Dec 28;6(12):e29733. doi: 10.1371/journal.pone.0029733

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Ferlin et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A. TUNEL analysis on human osteoblasts stimulated overnight with INSL3 at 1 nM, 10 nM and 100 nM. Positive control is obtained treating cells with DNase. Green fluorescence indicates DNA fragmentation. Nuclei were counterstained with DAPI (Blue). B. Quantitative RT-PCR for ALP in human osteoblasts stimulated with INSL3 at different concentrations. Quantification was normalized to the expression of the housekeeping gene β₂-microglobulin on control (no stimulation). Data are shown as the mean ± standard deviation (SD) of the mean of three different experiments performed in triplicate. C. Immunoenzymatic assay of ALP production in human osteoblasts after a 24 hour stimulation with INSL3 at different concentration. Control is obtained without stimulation. Data are shown as the mean ± standard deviation (SD) of the mean of three different experiments performed in triplicate. * P<0.05 vs control.