Journal: Journal of Animal Science

Article Title: Bta-miR-34b regulates milk fat biosynthesis by targeting mRNA decapping enzyme 1A (DCP1A) in cultured bovine mammary epithelial cells 1

doi: 10.1093/jas/skz230

Figure Lengend Snippet: Primers for mRNA quantitative real-time PCR

Article Snippet: The antibodies were used as follows: GAPDH antibody (1:10,000, Novus), anti-DCP1A antibody (1:1,000, Boster), and goat anti-IgG H&L (HRP; 1:2,000, Abcam).

Techniques: Sequencing

Journal: Journal of Animal Science

Article Title: Bta-miR-34b regulates milk fat biosynthesis by targeting mRNA decapping enzyme 1A (DCP1A) in cultured bovine mammary epithelial cells 1

doi: 10.1093/jas/skz230

Figure Lengend Snippet: (A and B) Relative expression levels of DCP1A mRNA in bovine mammary epithelial cells (BMEC) transfected with the miR-34b mimic (50 nM) and miR-34b inhibitor (200 nM) and their respective negative controls for 48 h. (C) Relative expression levels of DCP1A protein in bovine mammary epithelial cells (BMEC) transfected with the miR-34b mimic (50 nM) and miR-34b inhibitor (200 nM) and their respective negative controls for 48 h. (D) Sequence alignment of miR-34b and the 3′ untranslated region (UTR) of DCP1A using TargetScan (http://www.targetscan.org; accessed 6 May 2016) and miRBase (http://www.mirbase.org; accessed 6 May 2016) algorithms. (E) The 293T cells were cotransfected with the miR-34b mimic and a luciferase reporter containing a fragment of the DCP1A 3′ UTR harboring either the miR-34b binding site (DCP1A–3′ UTR wild-type [WT]) or a mutant (DCP1A–3′ UTR mutation type [MUTA]). Values are presented as means ± SEM; **P < 0.01; ***P < 0.001. miR = microRNA; NC = negative control; Rluc = Renilla luciferase; fluc = firefly luciferase.

Article Snippet: The antibodies were used as follows: GAPDH antibody (1:10,000, Novus), anti-DCP1A antibody (1:1,000, Boster), and goat anti-IgG H&L (HRP; 1:2,000, Abcam).

Techniques: Expressing, Transfection, Sequencing, Luciferase, Binding Assay, Mutagenesis, Negative Control

Journal: Journal of Animal Science

Article Title: Bta-miR-34b regulates milk fat biosynthesis by targeting mRNA decapping enzyme 1A (DCP1A) in cultured bovine mammary epithelial cells 1

doi: 10.1093/jas/skz230

Figure Lengend Snippet: (A) Validation of DCP1A knockdown in bovine mammary epithelial cells (BMEC) after the BMEC were transfected with DCP1A small interfering RNA (siRNA; 50 nM) and the control. (B and C) Changes in triacylglycerol and lipid droplet size in BMEC changed by DCP1A siRNA (50 nM). (D) The relative expression of lipid metabolism–related genes. Bovine mammary epithelial cells were transfected with DCP1A siRNA (50 nM) and the control for 48 h. Values are presented as means ± SEM; *P < 0.05; **P < 0.01; ***P < 0.001. NC = negative control; si-DCP1A = the siRNA of DCP1A.

Article Snippet: The antibodies were used as follows: GAPDH antibody (1:10,000, Novus), anti-DCP1A antibody (1:1,000, Boster), and goat anti-IgG H&L (HRP; 1:2,000, Abcam).

Techniques: Transfection, Small Interfering RNA, Expressing, Negative Control