Fig. 1: ASncmtRNA knockdown (ASK) induces inhibition of cell proliferation and apoptotic cell death of MDA-MB-231 breast cancer cells. | Cell Death & Disease

Fig. 1: ASncmtRNA knockdown (ASK) induces inhibition of cell proliferation and apoptotic cell death of MDA-MB-231 breast cancer cells.

From: Mitochondrial ncRNA targeting induces cell cycle arrest and tumor growth inhibition of MDA-MB-231 breast cancer cells through reduction of key cell cycle progression factors

Fig. 1

MDA-MB-231 cells were transfected with 200 nM Andes-1537 (1537) or ASO-C or left untreated (NT). a Knockdown of ASncmtRNA-1 and −2 at 24 h post-transfection was corroborated by RT-PCR, using 16 S and 18 S rRNA as controls. Numbers on the left represent size in bp of MW standards. b Viability was determined by MTT assay at 24, 48, and 72 h post-transfection. c At 48 h post-transfection, Andes-1537 induced over 60% death, determined by Trypan blue (Tb) exclusion assay, compared to 3–5% in controls (two-tailed Student’s t-test; Mean ± S.E.M. ****p < 0.0001; ASO-C vs. Andes-1537; n = 3). d Representative image of fluorescent TUNEL assay for determination of apoptosis, at 48 h post-transfection. Bars = 50 µm. e Ten fields per sample of the experiment shown in d were photographed and quantified. The graph shows % of TUNEL-positive cells (two-tailed Student’s t-test; Mean ± S.E.M. ****p < 0.0001; ASO-C vs. Andes-1537; n = 3). f Apoptosis was further confirmed at 48 h post-transfection by Annexin V-binding and co-staining with PI and analyzed by flow cytometry. g A triplicate analysis of the experiment in f shows increased Annexin V binding in Andes-1537-treated cells, compared to controls (two-tailed Student’s t-test; Mean ± S.E.M.****p < 0.0001; ASO-C vs. Andes-1537; n = 3)

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