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. 2018 Oct 22;8:15572. doi: 10.1038/s41598-018-33629-y

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Knockdown of BLAT1 Expression Increased Apoptosis of BLBC cell lines. (A) Two different antisense oligonucleotides (ASO) targeting exon 2 of BLAT1 (BLAT1a and 1b, respectively) are indicated by red bars. (B,C) qRT-PCR data showed an efficient knockdown of BLAT1 expression by BLAT1a and 1b ASOs in MDA-MB-468 (B) and HCC-1569 cells (C). (D, F) BLAT1 knockdown induced significant apoptotic response in MDA-MB-468 (D) and HCC-1569 cells (F) determined by caspase 3/7 activity. (E,G) Total apoptosis was shown to be increased by flow cytometry of Annexin V/PI stained cells 3 days after ASO knockdown in MDA-MB-468 (E) and HCC-1569 cells (G). Error bars, SE (***p < 0.001 vs. control-ASO treated cells). (H) Fifty micrograms of protein were subjected to mitochondrial OXPHOS complex I activity, which is determined by the oxidation of NADH to NAD+ and the simultaneous increase in absorbance at OD 450 nm. No significant difference in the activity was observed among MDA-MB-468 cells treated with control, BLAT1a or BLAT1b ASO. (I) Western blot analysis showed increased expression of γ-H2AX in BLAT1a or BLAT1b ASO-treated cells, compared to the control (Con). β-Actin was used as a loading control.