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. 2016 Sep 30;45(3):e12. doi: 10.1093/nar/gkw883

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — CRISPRi and siPOOL- / ASO-mediated knockdown of LOC389641, MNX1-AS1 and HOTAIR. (A–C) HLE cells expressing either dCas9 or dCas9-KRAB were transduced with either control sgRNA or one of five indicated sgRNAs targeting LOC389641. RT-qPCR results for LOC389641 (A) and TNFRSF10A (B) normalized to cyclophilin A and control sgRNA. Error bars represent SEM (n = 4). *P < 0.05; **P < 0.01; ***P < 0.001 compared to dCas9 / dCas9-KRAB + control sgRNA, unpaired two-sided t-test. (C) Western blot results for TNFRSF10A. VINCULIN was used as loading control. (D) HLE cells were transfected with either siPOOL control or siPOOL LOC389641. RT-qPCR results for TNFRSF10A and LOC389641 normalized to cyclophilin A and siPOOL control. Error bars represent SEM (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001 compared to control siPOOL, unpaired two-sided t-test. Western blot results for TNFRSF10A. VINCULIN was used as loading control. (E) NCI-H460 cells were transfected with either a control ASO or two independent ASOs against MNX1-AS1. Also, NCI-H460 cells expressing dCas9-KRAB were transduced with either control sgRNA or one of the three indicated sgRNAs targeting MNX1-AS1. RT-qPCR results for MNX1 and MNX1-AS1 normalized to cyclophilin A and control ASO / control sgRNA. Error bars represent SEM (n = 4). *P < 0.05; **P < 0.01; ***P < 0.001 compared to control ASO / sgRNA, unpaired two-sided t-test. (F) Hela cells were transfected with either siPOOL control or siPOOL HOTAIR. Also, Hela cells were transduced with lenti dCas9-KRAB-PURO iv sgRNA containing either control sgRNA or one of the three indicated sgRNAs targeting HOTAIR. RT-qPCR results for HOXC11 and HOTAIR normalized to cyclophilin A and siPOOL Control or control sgRNA. Error bars represent SEM (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001 compared to control ASO / sgRNA, unpaired two-sided t-test.