#Rfx 150 generator#

A.U., arbitrary unit CDS, coding sequence FC, fold change miRNA, microRNA RT-qPCR, reverse transcription quantitative PCR UTR, untranslated region WT, wild-type. (E) Same as in (D), except that the 4 putative miR-150 binding sites identified in the PDAP1 3вЂІ UTR were mutated by site-directed mutagenesis. Luciferase reads were normalized to the renilla ones. (D) The 3вЂІ UTR of the indicated genes was cloned in a dual-luciferase reporter vector and transfected into HEK293 cells together with either a miR-150 mimic or a control oligonucleotide. Twenty-four hours or 48 hours after transfection, the expression of the indicated genes was measured by RT-qPCR. (C) Activated memory cells were transfected with miR-150 mimic or control oligonucleotide. Both the 3вЂІ UTRs and the CDS were manually searched for the presence of at least a 6-mer miR-150 or miR-146a binding site. (B) The list of genes obtained from (A) was intersected with several prediction databases using miRWalk 2.0. Genes in red were considered significantly differentially expressed when log 2FC в‰Ґ 0.6, в€’log 10 p-value в‰Ґ 2.

(A) Volcano plot of differentially expressed genes between the indicated miRNA mimic and control oligonucleotides. CFSE, carboxyfluorescein succinimidyl ester miRNA, microRNA RT-qPCR, reverse transcription quantitative PCR. Data in the bar graph were normalized to the overall baseline signal on day 0, prior to stimulation, to compensate from experimental differences in basal CFSE loading. The extent of cell proliferation was measured 3 days after activation. (C) Freshly isolated memory T lymphocytes were loaded with CFSE, transfected with either a miR-150 mimic or a control oligonucleotide, and activated with anti-CD3 and anti-CD28 antibodies. MiRNA expression was measured by RT-qPCR, and data are expressed as 2 в€’О”Ct. (B) Total RNA was extracted from the indicated T-cell subsets freshly isolated from peripheral blood. The most highly expressed miRNAs are shown, and data are expressed as percentage of normalized counts over the total. (A) Total RNA was extracted from freshly isolated CD4 + naive, T CM, and T EM T-cell subsets, and miRNA expression was measured by NanoString SPRINT profiling.

Overall, using an integrated approach involving quantitative analysis, unbiased genomics, and genome editing, we identified RFX factors, miR-150, and the PDAP1 RBP as the components of a regulatory axis that restrains proliferation of primary human T lymphocytes. PDAP1 acted as an RNA-binding protein (RBP), and its CRISPR/Cas-9-mediated deletion revealed that it prominently contributed to the regulation of T-cell proliferation. By performing miRNA pull-down and sequencing experiments, we identified PDGFA-associated protein 1 (PDAP1) as one main target of miR-150 in human T lymphocytes. Constitutive MIR150 gene expression required the RFX family of transcription factors, and its activation-induced down-regulation was linked to their reduced expression. We found that in primary human memory T helper lymphocytes, miR-150 was the most abundantly expressed miRNA, and its expression decreased drastically upon activation, suggesting regulatory roles. However, what are the mRNA targets regulated by miRNAs and how miRNAs are transcriptionally regulated themselves remain for the most part unknown. Ransburg RansFlex RFXW Electrostatic Waterborne Direct ChargeWithin the immune system, microRNAs (miRNAs) exert key regulatory functions. this new feature allows the operator to keep the line moving, increasing production. The new вЂњQuick Fan AdjustвЂќ button allows operators to easilygo from a large to a small fan pattern by simply depressing the button located on the side of the gun. The RFQ and RFQX both offer superior finish quality surpasses competitorвЂ™s larger, higher voltage applicators in small compact size.

#Rfx 150 generator#

The RansFlex Quick Fan Adjust applicator, is an air atomizing electrostatic applicator powered by an onboard generator that provides best in class atomization in a lightweight, user friendly applicator. These advancements provide a compact applicator with the equivalent вЂ“ or better вЂ“ performance and transfer efficiency of higher voltage competitive guns

Smaller, Lighter weight RansFlex RX and RFX applicators feature enhanced, proprietary cascade and atomization technology. Advanced RansFlex Finishing Applicators Integrate The Best of Both Worlds RansburgВ® Electrostatics With Legendary DeVilbiss Atomization Technology.