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. 2017 Jul 14;7:5462. doi: 10.1038/s41598-017-05561-0

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© The Author(s) 2017

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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Northern blot analysis of miRNA expression in Chlamydomonas cells grown under the denoted trophic conditions. (A) Small RNAs were detected with probes specific for the indicated miRNAs. The same filters were reprobed with the U6 small nuclear RNA sequence as a control for lane loading. CC-124 wild type strain; Maa7-IR44s, CC-124 containing a transgene expressing FLAG-tagged AGO3; Mut-20, TSN1 deletion mutant, in the Maa7-IR44s background, defective in sRNA biogenesis¹⁴. (B) Relative miRNA levels in the indicated strains under the different trophic conditions. Values shown are the average of two independent experiments and are normalized to those of the Maa7-IR44s strain grown photoautotrophically in nutrient replete minimal medium (HS). For c20399 and c19166, values are normalized to those of Maa7-IR44s grown mixotrophically in acetate containing medium (TAP). The relative standard deviation, as percentage of the mean, was in no case higher than 28.3%. Data corresponds to phosphorimager measurements of sRNA signals on northern blots (gray bars) or normalized read counts from the AGO3-associated sRNA libraries (white bars).