Dataset Information

Matrix Remodelling during Lymph Node Expansion

ABSTRACT: In this study we focussed our investigations on ECM remodelling by FRCs during lymph node (LN) expansion, and the interconnection between the cellular and ECM components of the conduit network. We demonstrate a loss of ECM components of the conduit during acute LN expansion

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: Lukas Krasny

LAB HEAD: Paul Huang

PROVIDER: PXD015816 | Pride | 2021-05-11

REPOSITORIES: Pride

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Publications

Fibroblastic Reticular Cells Control Conduit Matrix Deposition during Lymph Node Expansion.

Martinez Victor G VG Pankova Valeriya V Krasny Lukas L Singh Tanya T Makris Spyridon S White Ian J IJ Benjamin Agnesska C AC Dertschnig Simone S Horsnell Harry L HL Kriston-Vizi Janos J Burden Jemima J JJ Huang Paul H PH Tape Christopher J CJ Acton Sophie E SE

Cell reports 20191101 9

Lymph nodes (LNs) act as filters, constantly sampling peripheral cues. This is facilitated by the conduit network, a tubular structure of aligned extracellular matrix (ECM) fibrils ensheathed by fibroblastic reticular cells (FRCs). LNs undergo rapid 3- to 5-fold expansion during adaptive immune responses, but these ECM-rich structures are not permanently damaged. Whether conduit flow or filtering function is affected during LN expansion is unknown. Here, we show that conduits are partially disru ...[more]

PMID: 31775047

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Project description:Treatment of the seeds. The Solanum lycopersicum var. Money Maker wild type (MM) and fri1-1 mutant seeds were incubated on distilled-water saturated cotton wool in clear plastic boxes wrapped in black plastic sheets in darkness during 3 hours at 25°C. After that, the seeds were treated as follows: a) 24 hr of continuous irradiation with far-red light, FRc; b) 24 hr of FRc follow by a 10 min red light pulse (Rp), FRc+Rp; and c) complete darkness. After the light treatments, the seeds were incubated in darkness (25°C) until sampling. Red light (35umol/m2sec) was provided by 40W fluorescent lamps (Phillips 40/15) in combination with a red translucid acrylic. FRc light (100 umol/m2sec) was provided by a set of 150W incandescent internal reflector lamps (Phillips) in combination with a red acetate filter, six 2mm thick blue acrylic filters Paolini 2031 and 10cm water filter. Total RNA isolation. The seeds were grinded in liquid nitrogen and extracted with two volumes of extraction buffer (1% SDS, 6% p-aminosalycilic acid, 1% NaCl, 6% water-saturated phenol and 2% 2-mercaptoethanol) and two volumes of acid phenol:chloroform solution. The aqueous phase was sequentially extracted with one volume of acid phenol:chloroform solution and one volume of chloroform. The nucleic acids in the aqueous phase were precipitated with 0.15 volumes of 3M NaCl and 2.5 volumes of 100% ethanol at –80°C for 20 minutes. After centrifugation, the pellet was washed with 70% ethanol, resuspended in RNase-free water and precipitated with one volume of 4M LiCl over night at 4°C. The LiCl was removed after centrifugation and the pellet was rinsed with 2M LiCl and resuspended in RNase-free water. The total RNA was treated with RNase-free DNase (Promega) and cleaned up with mini spin columns (RNeasy Plant Mini Kit, Qiagen). The mRNA was amplificated with Message Amp II aRNA Amplification Kit (Ambion) and concentrated by vacumm centrifugation. Keywords: Direct comparison 21 hybs total

Dataset Information

Matrix Remodelling during Lymph Node Expansion

Publications

Fibroblastic Reticular Cells Control Conduit Matrix Deposition during Lymph Node Expansion.

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