Figure 3

Isc Qu cotreatment prevents low-dose Dox induced senescence and SASP in MCF7 cells. Confluent MCF7 cells were treated with Isc Qu (85 µg/mL) and Dox (50 nM) alone or in combination for 72 hours and were subjected to immunofluorescence, FACS analyses and western blot. Immunofluorescent labeling of (A) p21 and (B) p53 protein expression, as well as (C) FACS analysis and immunobloting. Cell nuclei are labeled with DAPI (blue). (D) IL-6 protein levels in the supernatant of cultured MCF7 cells upon 72 h treatment with Isc Qu (85 µg/mL) and Dox (50 nM) alone or in combination, were measured using an ELISA. IL-6 activity was converted to pg according to the manufacturer’s instructions (Affymetrix, eBiosciences, CA, USA). (E) Protein expression of CCL-2 in treated MCF7 cells was determined by flow cytometry (left) and presented as MFI- the mean fluorescence intensity (right). Results are presented as the mean ± SEM of three independent experiments. Asterisks denote statistical significance compared to control cells (***p < 0.001). Representative photos and histogram for at least three independent experiments performed in triplicate are shown.