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Proteomics analysis in kidney membrane fraction of BDL- and sham-operated mice

ABSTRACT: Kidney tissues were collected from mice 2 weeks after BDL and sham operations. Membrane fractions of the kidney were prepared. After sample preparation, the proteome analysis was performed.

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Hiroshi Arakawa

PROVIDER: PXD036555 | JPOST Repository | Fri Sep 08 00:00:00 BST 2023

REPOSITORIES: jPOST

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			Action	DRS
	1_sham_mouse_kidney-1_20190914_20191003.raw	Raw
	2_sham_mouse_kidney-2_20190914_20191003.raw	Raw
	3_sham_mouse_kidney-3_20190914_20191003.raw	Raw
	4_sham_mouse_kidney-4_20190914_20191003.raw	Raw
	5_BDL_mouse_kidney-1_20190914_20191003.raw	Raw

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Project description:Aim and experimental set-up: Sequencing of small RNA from total RNA fractions. The aim of this experiment was to compare profiles of miRNA expression between the following genetic backgrounds: Col-0 (wild type), era1-2 (farnesyl transferase mutant), j2-2/j3-2 + pJ3:J3 (j2/j3 double knockout expressing transgenic J3 under the control of the endogenous J3 promoter), j2-2/j3-2 + pJ3:J3C417S (j2/j3 double knockout expressing transgenic J3 mutated in the farnesylation site (C417S) under the control of the endogenous J3 promoter). Seedlings were grown under sterile conditions for 16 days. Two biological replicates of each genotype were harvested, and total RNA was prepared by Trizol extraction. Small RNA libraries were constructed using NEBNext Small RNA Library Prep Set and sequenced on an Illumina platform. Sequencing of small RNA bound to AGO1 in membrane fractions The aim of this experiment was to characterize AGO1-bound small RNAs in membrane fractions, and to answer two specific questions: Can miRNAs be identified whose association with membrane-bound AGO1 is different between the following four genotypes: Col-0 (wild type), era1-2 (farnesyl transferase mutant), j2-2/j3-2 + pJ3:J3 (j2/j3 double knockout expressing transgenic J3 under the control of the endogenous J3 promoter), j2-2/j3-2 + pJ3:J3C417S (j2/j3 double knockout expressing transgenic J3 mutated in the farnesylation site (C417S) under the control of the endogenous J3 promoter) Is the ratio between reads matching miRNA and miRNA* different between the above four genotypes? Seedlings were grown under sterile conditions for 16 days. Two biological replicates of each genotype were harvested. Microsomes were prepared from 2g of seedling tissue, solubilized by 1% deoxycholate and AGO1 was immunoprecipitated with specific antibodies (Agrisera). Immunopurified AGO1 was eluted from Protein A sepharose beads by competitive elution with antigenic AGO1 peptide. Small RNA was extracted by Trizol extraction from eluted AGO1. Small RNA libraries were constructed using NEBNext Small RNA Library Prep Set and sequenced on an Illumina platform.

Dataset Information

Proteomics analysis in kidney membrane fraction of BDL- and sham-operated mice

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