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. 2018 Dec 5;8:17665. doi: 10.1038/s41598-018-35816-3

Figure 2.

Figure 2

Lack of cell cytotoxicity by HelixComplex treatment. In (A) fibroblasts were exposed to increasing doses of HelixComplex for analysis of cell viability (from 4 µg/ml to 400 µg/ml). Left panel: cell viability, examined by MTT colorimetric assays, was calculated at 24 hours as percentage with respect to the untreated cultures (set to 100%). Right panel: cell number was monitored over time for up to 72 hours, starting from a high density cell culture. DMSO (10%) was used as positive control of cell death. In (B) apoptosis was evaluated on fibroblasts treated for 48 hours with a high dose of HelixComplex (400 µg/ml) and calculated as percentage of Annexin V/PI double positive cells on the total population for each treatment. Representative plots of apoptotic cells analyzed by flow-cytometry are shown. In (C) fibroblasts were cultured for 48 hours in medium with 2% serum with or without 400 µg/ml HelixComplex. Medium with 10% serum was used in the untreated control (Untreated). Left panel: representative immunofluorescence images of actin organization (red staining). Nuclei were colored with DAPI (blue staining). Right panel: cell surface was measured and reported in arbitrary units (AU). In (A–C) data are reported as the mean ± SD of results from at least three independent experiments. The asterisk indicates p < 0.05 respect to untreated cultures. HC: HelixComplex.