Anti-Biotin Polyclonal antibody, PE

Pecan (Carya illinoinensis) is a widely consumed edible woody oil species that is rich in unsaturated fatty acids (FAs) that are beneficial to human health. However, the genes and mechanisms regulating seed oil biosynthesis in pecan are not well understood. Here, we analyzed the expression patterns of genes involved in seed oil biosynthesis in two different varieties of pecan with distinct fruit maturation schedules and oil contents. We cloned theC. illinoinensis WRINKLED 1(CiWRI1) gene, a homolog ofArabidopsisWRINKLED1(AtWRI1), which plays a key role in FA synthesis. OverexpressingCiWRI1restored lipid synthesis in theArabidopsiswri1-1mutant and rescued other phenotypic defects such as plant height, root length, and germination rate, suggesting thatCiWRI1is an ortholog of theAtWRI1and is involved in the regulation of FA synthesis. To investigate the mechanism ofCiWRI1regulation, we clonedC. illinoinensis BIOTIN CARBOXYL CARRIER PROTEIN ISOFORM2(CiBCCP2) and determined that the CiWRI1 protein directly binds to an ASML1/WRI1 (AW)-box motif in theCiBCCP2gene promoter and thereby activates its transcription.CiBCCP2overexpression partly rescued the phenotypic defects of thewri1-1mutant, indicating that it is directly regulated by CiWRI1. Thus, de novo FA biosynthesis in seed is conserved across plant species; moreover, CiWRI1 regulates oil synthesis by directly controllingCiBCCP2expression. These findings present novel potential targets for molecular-marker-assisted breeding of this commercially important plant.

Abdominal aortic aneurysm (AAA) is characterized as dilation of the aortic wall. Dysregulation of vascular smooth muscle cells (VSMCs) can contribute to the development of this phenotype. Circular RNAs and microRNAs (miRNAs) can regulate the proliferation and apoptosis of VSMCs. This present study aimed to identify the mechanisms of action behind the regulation of cerebellar degeneration-related protein 1 antisense RNA (CDR1as)/miRNA (miR)-7 in VSMCs. The expression levels of miR-7 were upregulated, whereas the levels of CDR1as and cytoskeleton-associated protein 4 (CKAP4) were downregulated in aortic specimens obtained from 10 patients who underwent surgery for AAA compared with aortic specimens from 10 control patients who underwent coronary artery bypass surgery. The molecular mechanism of action of CDR1as/miR-7 was investigated in primary VSMCs. The results of Cell Counting kit-8 and cell growth curve assays revealed that overexpression of CDR1as and knockdown of miR-7, increased VSMC proliferation, whereas knockdown of CDR1as and overexpression of miR-7 suppressed VSMC proliferation. In addition, overexpression of CDR1as and knockdown of miR-7, suppressed apoptosis in VSMCs, indicated by the decreased levels of reactive oxygen species (ROS) and lactate dehydrogenase (LDH) activity, whereas knockdown of CDR1as and overexpression of miR-7 exhibited the opposite effects. The results of luciferase reporter and biotin pull-down assays confirmed that CDR1as directly bound to miR-7 and suppressed its expression. Additionally, the CDR1as-induced proliferation and suppressed apoptosis was reversed by the overexpression of miR-7. Furthermore, luciferase reporter, reverse transcription-quantitative PCR and western blot assays revealed that miR-7 directly targeted CKAP4 and suppressed its expression. Additionally, the miR-7-suppressed proliferation and increased ROS and LDH activity were reversed by the overexpression of CKAP4. CDR1as also decreased caspase 3/7 activity, which was reversed by miR-7 mimics. miR-7 increased the activity of caspase 3/7, which was again reversed by the overexpression of CKAP4. Therefore, CDR1as, miR-7 and CKAP4 may act in the same pathway to regulate VSMC proliferation and apoptosis.