Fig 1: Species specific expression of MAOA in adipocytes.(A) Dot plots of indicated genes generated using data from Emont et al., 2022, available here (B) RT-PCR and western blotting at indicated days following induction of differentiation in human adipocytes. Samples were prepared from two independent wells and subjected to western blotting with an antibody to MAOA that recognizes the mouse and human proteins (CellSignaling #75330). The intensity of the single detected band was quantified, and values plotted in the right Y axis. RT-PCR results are plotted in the left Y axis. Symbols represent the fold difference over the lowest value in the dataset and bars the mean and SEM of n=4 independent cultures. (C) Representative images of human and mouse subcutaneous adipose tissue stained with antibodies to MAOA (CellSignaling #75330 for mouse, CellSignaling #73030 for human), and PLIN1, and counterstained with DAPI. Bars = 100 um. (D) Confocal images of human adipose tissue stained with antibodies to PLIN1, MAOA (CellSignaling #73030), and mtHSP70 and counterstained with DAPI. Projection of 15 images taken at 0.5 µm intervals (bar=10 um) and indicated segment in 3D projections illustrating localization of nucleus, mitochondria and MAOA to the region between the lipid droplet and the cell periphery (bars = 3 um). (E) WIM and vicinal mouse adipose tissue, stained for PLIN1, MAOA (CellSignaling #75330), and DAPI, with red dotted line indicating boundary between hybrid tissue and endogenous adipose tissue. Bar = 100um. (F) Immune cell showing staining for MAOA. Bar = 10 µm. (G) Intensity of MAOA (left panel) and PLIN1 (right panel) relative to DAPI in 5 independently analyzed regions from each of n=4 WIM and n=4 TIM thin sections, as described in Materials and methods. Statistical significance of the differences was assessed using two-tailed student t-tests, and exact values are shown. (H) Experimental timeline for exposure on mice harboring WIM to clorgyline or cold. (I) Weight of mice at 7- and 8 weeks (J) Weights of dissected tissues at 8 weeks. (K) RT-PCR for human UCP1. (L) RT-PCR for human ADIPOQ. For (K and L), bars represent the mean and S.E.M and symbols correspond to each independent implant (n=7–8 implants). M. RT-PCR for mouse Ucp1 in excised inguinal adipose tissue (n=7–8 fat pads). Bars represent the mean and S.E.M and symbols correspond to each independent fat pad. (I-M) Statistical significance of the differences was assessed using one-way ANOVA with Dunett’s multiple comparison correction, and exact values are shown. Figure 6—source data 1.Uncropped, labeled western blot.