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. 2021 Nov 30;233(3):1257–1273. doi: 10.1111/nph.17861

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© 2021 The Authors. New Phytologist © 2021 New Phytologist Foundation

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Fig. 1 — miR397 modulates expression of fluorescently tagged CsiLAC4 and CsiLAC17 in tobacco leaf via miRNA‐mediated cleavage of mRNA. (a) miRNA blot showing generation of mature miR397 by transient expression of pre‐miR397 in tobacco leaves. Total RNAs were extracted 48 h postinoculation. (b) 5′‐RACE PCR of CsiLAC4 and CsiLAC17 from tobacco leaves coexpressing miR397 and CsiLAC4‐DsRed/CsiLAC17‐DsRed. Arrowheads indicate expected fragments supporting the miRNA‐mediated cleavage. (c) Site‐directed mutations of CsiLAC4 and CsiLAC17 in complementary sites of miR397. Mutated nucleotides are indicated in red. (d) miR397 represses expression of DsRed‐tagged CsiLAC4 and CsiLAC17 rather than their corresponding mutants in the tobacco epidermis. Confocal micrographs were obtained 48 h posttransfection. GFP was used as an indicator of successful transfection.