Extended Data Fig. 8: B cell-derived acetylcholine in mice with atherosclerosis.

a. Cd19^CreChat^fl/fl mice or their Chat^fl/fl littermates were intravenously injected with an adeno-associated virus 2/8 encoding hPCSK9 and fed a high-fat diet (HFD) for 12 weeks. Serum cholesterol levels after 12 weeks of HDF. Two independent experiments, n = 6 mice per Chat^fl/fl group and n = 8 per Cd19^CreChat^fl/fl group, two-tailed Mann-Whitney test. b. Aortic roots were harvested from Chat^GFP mice with atherosclerosis for flow cytometry and histology analysis. GFP⁺ B cell abundance in the femur and aorta of Chat^GFP mice assessed by flow cytometry. P = 0.0038 absolute number data and P = 0.0094 for relative frequency data. n = 4 mice per group, two-tailed Student’s t test. c. Immunofluorescence image of the aortic roots in Chat^GFP mice (green). Sections were stained with B220 (red) and DAPI (blue). The experiment was performed with 4 independent mice, which all showed a similar result. Scale bar, 200 µm. d. Higher magnification images of areas indicated in e. Arrow heads indicate Chat^GFP-positive nerve fiber, arrows indicate B220⁺ B cells. The experiment was performed with 4 independent mice, which all showed a similar result. Scale bar in panel 1, 10 µm, all other panels 20 µm. Data are mean ± SEM with individual values for data distribution, **P < 0.01.