Data independent acquisition mass spectrometry of isolated glomeruli from mutant mice lacking the nuclear export signal of p62 and WT

＜Project description＞
Data independent acquisition (DIA) mass spectrometry of Isolated glomeruli from homozygous deletion of the nuclear export signal of p62 (dNES) and WT mice (n=3 each).

＜Sample processing＞
Formalin fixed paraffin embedded (FFPE) tissues were prepared in general method and kidney samples were cut into 5µm-thick sections on the membrane slides (Leica Microsystems, Mannheim, Germany). Glomeruli were isolated via laser capture micro-dissection (0.6 mm2 in total).
The dissected glomeruli were collected separately into 40 µL of Tris EDTA buffer and 0.002% Zwittergent 3-16 detergent.　Solubilised proteins were further digested into peptides by 0.01ug/ul trypsin (Promega, Madison, WI, USA) overnight. Each sample was centrifuged at 15000 rpm for 30 minutes prior to measurement.

＜Mass spectrometry and data processing＞
Samples were measured by DIA-MSMS using NanoLC-MSMS system with Vanquish Neo and Orbitrap Exploris 240 (Thermo Scientific).
The measurement data were analyzed with DIA-NN 1.8.1.