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. 2020 Feb 6;10:2008. doi: 10.1038/s41598-020-58495-5

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© The Author(s) 2020

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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Functional analysis of MID1 in mitochondria. (A) qRT-PCR analysis of splicing efficiency of mitochondrial introns in mid1. The histograms display the ratio of the spliced transcripts to the unspliced transcripts in mid1 plants as compared with wild-type in a log₂ form. Each mean value is the average of three biological replicates, each of which undergoes triple repeats. s, spliced transcripts; un, unspliced transcripts. (B) RT-PCR analysis of mitochondrial transcripts splicing efficiency. u1 means nad2 transcript with intron 1.m means mature nad2 transcript. comp, MID1Pro:MID1-3 × Flag complemented line. (C) BN-PAGE analysis of mitochondrial protein complexes abundance in wild-type, mid1 mutants, mtsf-1 and complemented line. Protein marks are indicated in the left panel and the bands representing main protein complexes are listed in the right panel. (D) In-gel analysis of complex I activity in wild-type, mid1 mutants, mtsf1 and complemented line. comp, MID1Pro:MID1-3 × Flag complemented line. Transmission electron microscopy analysis of mitochondrial ultrastructure in leaf of 4-week-old WT (E) and mid1-1 (F). Bars = 0.5 μm. (G) Mitochondrial area of wild-type and mid1-1. Data are the mean ± SE, n = 140. Student’s t tests, *P < 0.05. (H) ATP content in the leaf of wild-type and mid1-1, Student’s t tests, **P < 0.01.