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. 2019 Mar 1;47(8):3937–3956. doi: 10.1093/nar/gkz128

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© The Author(s) 2019. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Effects of DR2 Alu RNA on differentiation and proliferation of PC-3 cells. (A) Northern blot of RNA harvested from PC-3 cells after 48 h treatment with or without 20 μM ML-60218 and hybridized with probes for U2 snRNA (pol II-transcribed control) and each strand of NANOG-Alu-Sx RNA. (B) Means of the relative changes in normalized expression of unprocessed pre-tRNA^Tyr and mRNAs encoding CK8, CK14, CK18, SYP, NSE, GRP78, CD55, CD59, CD63 and NANOG, as determined by RT-qPCR in 3–5 independent experiments, in PC-3 cells harvested 48 hrs after transfection with a 33nt synthetic RNA corresponding to part of the NANOG-Alu-Sx sequence. (C) Western blot of NANOG and actin protein expression in PC-3 cells 48 h after mock transfection (control) or transfection with a 33nt synthetic RNA corresponding to forward or reverse strands of part of the NANOG-Alu-Sx sequence. (D) Means of PC-3 cell numbers over 3 days following transfection with a 33nt synthetic RNA corresponding to part of the NANOG-Alu-Sx sequence, as indicated, in 4 independent experiments. (E) Means of the percentage of PC-3 cells staining positively for Ki67 proliferation marker 48 h after mock transfection or transfection with a 33nt synthetic RNA corresponding to part of the NANOG-Alu-Sx sequence or a 56nt synthetic RNA corresponding to part of the DR2 Alu consensus sequence. (F) Means of the relative changes in normalized expression of unprocessed pre-tRNA^Tyr and mRNAs encoding CK8, CK14, CK18, SYP, NSE, GRP78, CD55, CD59, CD63 and NANOG, as determined by RT-qPCR in three independent experiments, in PC-3 cells harvested 48 h after transfection with a 56nt synthetic RNA corresponding to part of the DR2 Alu consensus sequence. (G) Means of PC-3 cell numbers over 3 days following transfection with a 56nt synthetic RNA corresponding to part of the DR2 Alu consensus sequence, as indicated, in three independent experiments. * indicates P < 0.05 relative to control by t-test; ** indicates P < 0.01; *** indicates P < 0.005. Error bars represent S.E.M.