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. 2018 Jun 30;46(17):9148–9159. doi: 10.1093/nar/gky591

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© The Author(s) 2018. 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 Non-Commercial 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 5. — SMA1 is required for MIR transcription. (A) Pri-miRNA levels in Col-0 and sma1-1 detected by RT-qPCR. Pri-miRNA levels in sma1-1 were normalized to those of UBQ5 and compared with Col-0 (set as 1). (B) Histochemical GUS staining of five-day-old seedlings and inflorescences in Col-0 and sma1-1 harboring the pMIR167a::GUS transgene. Fifteen plants containing GUS were analyzed for each genotype and one image for each genotype is shown. (C) GUS transcript levels in Col-0 and sma1-1 harboring the pMIR167a::GUS transgene. GUS transcript levels detected by RT-qPCR were normalized to those of UBQ5 and compared with Col-0 (set as 1). (D) The occupancy of Pol II at MIR promoters in Col-0 and sma1-1 detected by ChIP followed by qPCR. The intergenic region between At2g17470 and At2g17460 (POL II C1) was used as a negative control. IP was performed using anti-RPB1 antibodies. (E) The association of GFP and GFP-SMA1 with MIR promoters in transgenic plants containing 35S::GFP or 35S::GFP-SMA1. The occupancy of GFP-SMA1 and GFP at MIR promoters was detected by qPCR following ChIP. The intergenic region between At2g17470 and At2g17460 (POL II C1) was used as a negative control. IP was performed using anti-GFP antibodies. Error bars (A–E) indicate standard deviations (SD) of three replicates. * in A and C: P < 0.05 with one-sample t-test. Means with different letters are significantly different determined by ANOVA (P < 0.05).