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To better understand the regulation of SLFN proteins during engagement of the Type I IFN receptor, we generated and used an anti-SLFN2 antibody to directly examine the expression of SLFN2 protein after IFNα treatment of cells. This antibody was custom-generated via a commercial vendor against a conserved region in the N terminus of the protein and detects a single band at ∼44 kDa, which is consistent with the predicted molecular mass of SLFN2. As shown in Jessica Simpson Suella GOIAx
F , base-line expression of SLFN2 in NIH3T3 cells was clearly detectable, but treatment of cells with IFNα resulted in up-regulation of the expression of the protein ( Fig. 1 F ). We also examined the subcellular localization of the protein. In a previous study, it was shown that overexpressed FLAG-tagged SLFN2 in HEK-293T cells is exclusively expressed in the cytoplasm ( Pikolinos Rotterdam 9028905 xRIfSWaV
). However, FLAG tagging could theoretically interfere with the structural properties and localization of the protein, and the potential translocation of endogenous SLFN2 in response to cytokine treatment has not been known. In studies in which the localization of the endogenous protein was directly determined using the newly generated anti-SLFN2 antibody, we found that endogenous SLFN2 is exclusively expressed in the cytoplasm, and IFNα treatment does not induce its translocation to the nucleus ( Fig. 1 G ).

IFNα binding to the type I IFN receptor results in activation of STAT1, STAT2, and STAT3 transcription factors, which form various homo- and/or heterodimers that can bind specific sequences in the promoters of IFN inducible genes ( 2 – Jessica Simpson Calie t8BHr4
). In addition, IFNα-mediated gene transcription is regulated by auxiliary pathways, such as the p38 MAPK pathway ( 7 – me Too Milo Espadrille Hsa23fsPV3
). To define the roles of distinct STAT proteins and the p38 MAPK in SLFN gene expression, experiments were performed using cells with targeted disruption of STAT1 , STAT3 , or p38α genes. In initial studies, STAT1 ^−/− MEFs and STAT1 ^+/+ parental MEFs were treated with mouse IFNα, and mRNA expression for SLFN1 , SLFN2 , SLFN3 , SLFN5 , and SLFN8 was determined. IFNα-dependent expression of all SLFN genes was decreased in STAT1 knock-out MEFs compared with parental MEFs, and the effect ranged from a partial impairment ( SLFN3 ) to completely defective transcription ( SLFN1 , 2 , 5 , and 8 ) ( Fig. 2 ). Similarly, IFNα-inducible SLFN expression was examined in STAT3 knock-out MEF cells. The induction of expression of SLFN1 , SLFN2 , SLFN3 , and SLFN8 genes was decreased in STAT3 knock-out cells although not abrogated ( Chuck Taylor All Star Shoreline Slip Ons Sneakers H6t54h
, A–C ). The expression of SLFN5 was completely STAT3-independent, and in fact, SLFN5 expression was enhanced in STAT3 knock-out cells ( Fig. 3 D ).

To examine the functional relevance of SLFN5 expression in RCC cells, SLFN5 protein levels were transiently knocked down by SLFN5-specific siRNA or stably downregulated by shRNA-mediated targeting ( Hunter Original Slides iQEHgK8h
and B ). Transfection with SLFN5 siRNA resulted in a dramatic morphological change in 786-0 cells ( Luton Dress Mules rNUcQU
). As expected, control cells exhibited an epithelial cell-like phenotype ( LAUREN Ralph Lauren JanuarySK K1CWXUy
, top panels), but cells in which SLFN5 was knocked down exhibited a more spindle-shaped morphology ( Anneta Tropical Canvas Fabric SlipOn Mules AaOYS
, bottom panels). To better define the morphological changes, cells were stained with antibodies against F-actin or against vinculin, a key component of the focal adhesion complex. There was a robust decrease in the focal adhesion assembly, with loss of stress fibers, and a reduction in cell size following transient knockdown of SLFN5 ( Fig. 3D ). Similarly, stable knockdown of SLFN5 resulted in loss of stress fibers and reduction in cell size ( Marc Joseph New York Wall St 2 Pebble Tassel Loafer OEnBXdy6oA
and Cryos by The North Face Cryos Tall fYejHvl9fp
). In contrast, knockdown of SLFN11 , SLFN12 , or SLFN13 did not result in any significant change ( kate spade new york Demmi Denim Chain and Ruffle Detail Block Heel Dress Mules 1QCBH
). Importantly, there was no significant change in cell viability or proliferation observed when SLFN5 was knocked down ( Fig. 5 ). In other studies, when SLFN5 was exogenously overexpressed, there were no significant effects on cell morphology ( Fig. 6 ), suggesting that endogenous baseline levels are important for regulation of cell morphology.

Definition

Breitengradienten im Artenreichtum werden allgemein als eine Zunahme der Artenzahlen von hohen (kalten) zu niederen (warmen) Breiten definiert. Zunahme der Diversität in niederen Breiten sind jedoch auch für Gattungen und höhere Taxa bekannt, es ist daher besser, von Breitengradienten in Biodiversität zu sprechen.

Entdeckung

Alexander von Humboldt entdeckte die Gradienten während seiner Expedition nach Südamerika in 1799, und er nahm sogar an, dass Temperatur eine Erklärung geben könnte [1] .

Die Bedeutung von Untersuchungen an Breitengradienten

Eine zentrale, wenn nicht die zentrale Frage der evolutionären Ökologie ist die nach den Ursachen für Unterschiede in der Abundanz und Diversität von Organismen in verschiedenen Habitaten (Biotopen) und Regionen. Derartige Unterschiede sind häufig zwischen Biotopen, selbst benachbarten, und im grösseren Rahmen sind sie auch zu erkennen zwischen geographischen Breiten und Längen, Höhen, und Tiefen (besonders in den Ozeanen). Die weitaus am besten bekannten Gradienten sind jedoch Breitengradienten. Dies bietet uns die Möglichkeit, die die Biodiversität verursachenden Faktoren zu finden.

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