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. 2019 Dec 27;45(2):543–555. doi: 10.3892/ijmm.2019.4443

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Copyright: © Li et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 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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Restoration of MED1 inhibits the metabolic action of miR-146a. (A and B) The protein levels (A) and relative mRNA levels (B, n=3) of MED1 in AML12 cells transfected with pcDNA3.1 or pcDNA-MED1. (C-E) The protein level of MED1 (C) and relative mRNA levels of miR-146a (D, n=3) and MED1 (E, n=3) in AML12 cells cotransfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1. (F and G) Representative images of Oil-red O staining (F) and cellular TG content analysis (G) of AML12 cells transfected with NC and pcDNA3.1, miR-146a mimics and pcDNA3.1 or miR-146a mimic and pcDNA-MED1. Scale bar, 100 µm. (H and I) The relative mtDNA copy numbers (H) and ATP levels (I) in AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 (n=3). (J) The relative mRNA expression level of nuclear respiratory factors (Nrf1 and Nrf2) and their downstream target genes (Tfam, Plormt and Tfb1m) in AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 (n=3). Restoration of MED1 inhibits the metabolic action of miR-146a. (K) Relative mRNA expression of oxidative phosphorylation pathway-related genes in the nucleus (Ndufs3, Sdhb, Sdhc and Cox5b) and in mitochondria (ND4L, CYTB, ND1, COII, and 12SrRNA) in AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 (n=3). (L) The relative mRNA expression level of nuclear respiratory factors and their downstream target genes in AML12 cells transfected with Scramble control or miR-146a inhibitor (n=3). (M) Relative mRNA expression of oxidative phosphorylation pathway-related genes in the nucleus and in mitochondria in AML12 cells transfected with Scramble control or miR-146a inhibitor (n=3). (N) Oxygen consumption of AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 were analyzed with a Seahorse XF24 extracellular flux analyser (n=3). (O) Basal respiration and respiratory capacity oxygen consumption rates were measured and calculated (n=3). Means ± SD are shown. ^*P<0.05; ^**P<0.01; ^***P<0.001 (vs. control group transfected with NC and pcDNA3.1), ^#P<0.05; ^##P<0.01; ^###P<0.001 (vs. the group transfected with miR-146a mimic and pcDNA3.1).

Restoration of MED1 inhibits the metabolic action of miR-146a. (A and B) The protein levels (A) and relative mRNA levels (B, n=3) of MED1 in AML12 cells transfected with pcDNA3.1 or pcDNA-MED1. (C-E) The protein level of MED1 (C) and relative mRNA levels of miR-146a (D, n=3) and MED1 (E, n=3) in AML12 cells cotransfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1. (F and G) Representative images of Oil-red O staining (F) and cellular TG content analysis (G) of AML12 cells transfected with NC and pcDNA3.1, miR-146a mimics and pcDNA3.1 or miR-146a mimic and pcDNA-MED1. Scale bar, 100 µm. (H and I) The relative mtDNA copy numbers (H) and ATP levels (I) in AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 (n=3). (J) The relative mRNA expression level of nuclear respiratory factors (Nrf1 and Nrf2) and their downstream target genes (Tfam, Plormt and Tfb1m) in AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 (n=3). Restoration of MED1 inhibits the metabolic action of miR-146a. (K) Relative mRNA expression of oxidative phosphorylation pathway-related genes in the nucleus (Ndufs3, Sdhb, Sdhc and Cox5b) and in mitochondria (ND4L, CYTB, ND1, COII, and 12SrRNA) in AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 (n=3). (L) The relative mRNA expression level of nuclear respiratory factors and their downstream target genes in AML12 cells transfected with Scramble control or miR-146a inhibitor (n=3). (M) Relative mRNA expression of oxidative phosphorylation pathway-related genes in the nucleus and in mitochondria in AML12 cells transfected with Scramble control or miR-146a inhibitor (n=3). (N) Oxygen consumption of AML12 cells co-transfected with NC and pcDNA3.1, miR-146a mimic and pcDNA3.1 or miR-146a mimic and pcDNA-MED1 were analyzed with a Seahorse XF24 extracellular flux analyser (n=3). (O) Basal respiration and respiratory capacity oxygen consumption rates were measured and calculated (n=3). Means ± SD are shown. ^*P<0.05; ^**P<0.01; ^***P<0.001 (vs. control group transfected with NC and pcDNA3.1), ^#P<0.05; ^##P<0.01; ^###P<0.001 (vs. the group transfected with miR-146a mimic and pcDNA3.1).