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. Author manuscript; available in PMC: 2023 Jul 1.
Published in final edited form as: Nature. 2022 Dec 7;613(7942):160–168. doi: 10.1038/s41586-022-05507-1

Extended Data Fig. 10 ∣. Conservation of CLSTN3B in humans.

Extended Data Fig. 10 ∣

a, Amino acid alignment of CLSTN3β from representative placental mammals. b, RNA-seq reads at the CLSTN3 locus in human cortex and adipose tissue. c, Microarray analysis of CLSTN3 expression in periadrenal adipose tissue from control patients, pheochromocytoma patients with unilocular fat (PheoUni), and pheochromocytoma patients with multilocular fat (PheoMulti) (n = 4). d, qPCR analysis of CLSTN3 exons 1/2, CLSTN3 exons 17/18, CLSTN3B, and UCP1 expression in human unilocular and multilocular adipose tissue (n = 26, 12). Unilocular group includes both control and PheoUni patients. e, RNAscope analysis of CLSTN3B expression in unilocular periadrenal adipose tissue from lean control, obese control, and pheochromocytoma (PheoUni) patients. Scale bar = 50 μm. f, Cloning of human CLSTN3B. g, Confocal microscopy of OA-treated (1 mM, overnight) HeLa cells transfected with human CLSTN3β-Flag and stained with LipidTOX 647. Scale bar = 5 μm. h, Amino acid alignment of mouse and human CLSTN3β (reference sequences). Putative hydrophobic hairpins, β-strand domain, and TM domain are colored red, blue, and purple, respectively. Underlined residues are altered in the patient from which human CLSTN3β was cloned for this study. Bar plots show mean ± SEM. Each point represents a biological replicate. Two-sided *P < 0.05, **P < 0.01, ***P < 0.001 by ordinary one-way ANOVA with Dunnett’s multiple comparisons test (c) or multiple t-tests with Holm-Sidak correction (d).