Supplementary Figure 3 from miR-34a Silences c-SRC to Attenuate Tumor Growth in Triple-Negative Breast Cancer

Figure S3, Related to Figure 4. A, c-SRC expression in normal (HFF and MCF-10A), luminal-A (MCF-7), LAR-TNBC (MDA-MB-453), and mesenchymal-TNBC (MDA-MB-231, BT-549, and Hs578T) cell lines. Expression was normalized to GAPDH and made relative to c-SRC expression in HFF lines. B-C, Clonogenic assays in Hs578T (B), and MDA-MB-436 (C) TNBC cells after miR-34a transfection and treatment with Dasatnib (left panels) and paclitaxel (right panel) or as a control HeLa cells after dasatinib treatment (C, right panel). D-E, Assessment of miR-34a target genes in MDA-MB-231 cells after 72 hours of dasatinib treatment (D), or miR-34a levels after 72 hours paclitaxel treatment (E). F, miR-34a promoter luciferase assays in MDA-MB-31 cells after dasatinib treatment. G-H, Spearman rank correlation analysis of SRC levels in cell lines described in Figure 1A (G), and in all breast cancer samples in the Metabric dataset (H). I, Comparable decreases in phospho-Tyr416 active c-SRC, non-phospho-Tyr527 c-SRC, and total c-SRC are observed in MDA-MB-231 cells transfected with 15nM miR-34a versus miR-Scr control, as determined by Western blot analysis. J, Schematic highlighting the miR-34a-c-SRC double-negative feedback loop present in MSL TNBC cells, which can be influenced by exogenous addition of miR-34a, or by dasatinib treatment. * Indicates p<0.05, as compared to control conditions.