Fig. 6: Sympathetic denervation of iLN compromises IL-33 release, dampens ILC2 activation and beiging of scWAT in response to cold challenge.

Eight-week-old male C57BL/6 N mice were subjected to bilateral sympathetic denervation of iLN by local injection of 6-OHDA (9 mg/ml in 0.15 M NaCl) or 0.15 M NaCl (Veh) directly into iLNs. Afterwards, mice were housed at 30 °C for 3 weeks followed by cold exposure (6 °C) or continued to be housed at 30 °C for another 2 days. a A macroscopic image showing the injection site of iLN embedded in scWAT. b LC/MS-based measurement of norepinephrine content in iLN and its surrounding inguinal scWAT (n = 5). c The mRNA expression of thermogenic genes in scWAT (n = 6). d UCP1 protein level in scWAT (top) and densitometric analysis for the relative abundance of UCP1 normalized with HSP90 (bottom) (n = 3). e Representative images of IHC staining for UCP1 in scWAT. Scale bar, 200 μm. f IL-33 protein level in iLN (left) and densitometric analysis for the relative abundance of IL-33 normalized with HSP90 (right) (n = 3). g ELISA analysis of IL-33 protein level in scWAT (n = 5). h, i Quantification of the percentage of IL-5- and IL-13- (h), and MetENK- (i) positive cells in activated ILC2s using flow cytometric analysis (n = 5). j Flow cytometric analysis for the proportion of ILC2s in lineage negative leukocytes in SVF. For Veh_30 °C, Veh_6 °C and 6-OHDA_6 °C (n = 6) or 6-OHDA_30 °C (n = 8). All samples are biologically independent replicates. Data are presented as mean ± SEM. Statistical data were assessed using the Mann–Whitney U test (c, g–i) or unpaired two-tailed Student’s t test (b, d, f). All the p values were two-sided. Source data are available as a Source Data file. kDa, relative molecular weight in kilodalton; NS not significant. See also Fig. S18.