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. 2021 Nov 13;10(11):3152. doi: 10.3390/cells10113152

Figure 2.

Figure 2

mRNA targets of miR 208a in cardiomyocytes incubated in diabetogenic conditions. (A) SV40 cardiomyocytes with normal (208+) and deficient (208−) miR 208a. First, 2 µg of total RNA from each sample was used to generate cDNA. RT-PCR cDNA products were used as templates for quantitative PCR (qPCR). All the qPCR assays were run in duplicate. RT-PCR products were separated on 1.5% agarose gels, while the bands were quantified and expressed relatively to the housekeeping mRNA U6. A Quick-Load® 100 bp DNA Ladder was used as a molecular weight marker. (B) Stanniocalcin 1 (STC1). (C) Mediator complex subunit 7 (MED7). (D) Sortin nexin 10 (SNX10). (E) Mitochondrial ribosomal protein S28 (MRPS28). (F) Mitochondrial contact site and cristae organizing system protein 1 (MINOS1). SV40 human cardiomyocytes were incubated in DMEM with 5 mM glucose (N), 25 mM glucose (high glucose, HG), or 20 µM bovine serum albumin (BSA)-bound palmitate (high palmitate, HP) for 24 h. First, 2 µg of total RNA from each sample was used to generate cDNA. RT-PCR cDNA products were used as templates for quantitative PCR (qPCR). All the qPCR assays were run in duplicate. RT-PCR products were separated on 1.5% agarose gels, while the bands were quantified and expressed relatively to the housekeeping mRNA β-actin. The alignment between miR 208a sequence and specific mRNAs, as well as the level of conservation of those sequences in different species, is shown in Supplementary Figure S1. Results are expressed as the mean ± SEM of three independent experiments. * p < 0.05.