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. 2022 Nov 25;13:7269. doi: 10.1038/s41467-022-34846-w

Fig. 6. Negr1 levels and secretion are reduced in the PVAT of ob/ob mice.

Fig. 6

a MS-determined relative abundances of Negr1 in the secretomes of wt and ob/ob AR and TH PVAT. N = 6 biological replicates per group, unpaired, two-tailed t-test, mean ± SEM is shown. b Western blot on wt TH PVAT lysate (LYS.) and supernatants (SN.) pools (N = 6 biological replicates per pool) stained for Negr1. c Representative immunofluorescence images (1 out of 2 replicates) showing Negr1 localisation in wt TH PVAT. Adipocytes are labelled by perilipin. Nuclei are counterstained with DAPI. Insets show higher magnification of white framed areas. Scale bar: 50 µm. d Western blot showing Negr1 and perilipin levels in stromal vascular fraction (SVF) and adipocytes obtained after digestion and fractionation of VI and SC WAT and BAT explants from wt mice (1 out of 2 replicates). e Western blot on primary murine sympathetic neurons lysates from 3 biological replicates stained for TYR.H. and Negr1. f Western blot on supernatants from TH (top) and AR (bottom) PVAT explants from wt mice incubated for 24 h in absence (DMSO) or presence of the wide spectrum MMP inhibitor GM6001 and stained for Negr1 and PAI1. N = 4 biological replicates per group. g and h Western blots showing Negr1 abundances in wt and ob/ob TH (g) and AR (h) PVAT lysates and relative quantifications. N = 5 biological replicates, unpaired two-tailed independent t-test (unequal distribution). Data are shown as mean ± SEM. AR aortic arch PVAT, TH thoracic PVAT, VI visceral AT, SC subcutaneous AT, BAT interscapular brown AT, TYR.H. tyrosine hydroxylase, Negr1 neuronal growth regulator 1, PAI1 plasminogen activator inhibitor-1, SCG superior cervical ganglia, M molecular weight marker. Source data are provided as a Source Data file.