Extended Data Fig. 5: CD45 (PTPRC) identification as a marker for SA/VA 3 + 4 positive adipocytes in human adipose tissue using different approaches.

A. Representative IHC images of hSAT and hVAT sections stained with anti-CD45 Ab, with and without counterstaining to highlight the positive signal from the adipocytes. scale bar = 20 μm. Shown are representative images of adipocytes from 6-8 different patients (from at least 2 independent staining experiments). B. panel of representative images showing lymph node section expressing the specific marker used as a positive control, no-primary antibody as a negative control, and the positive signal from within stained adipose tissue. The fluorescence emission spectra obtained from these slides are displayed in the graph. White squares indicate the areas where spectrum measurements were taken. Shown are results of a single experiment. C-D. Representative images of adipocytes displaying positive signal for CD45 taken from hSAT and hVAT of two different patients (each panel shows a different patient). scale bar = 20 μm. These 2 patients are representative images of adipocytes from 6-8 different patients (from at least 2 independent staining experiments). E. Image-stream flow cytometry analyses of isolated human adipose tissue cells. PTPRC protein (CD45, common leukocyte antigen) is shown in SVF cells, potentially macrophages, and CD45-negative adipocytes (based on round morphology, diffuse Bodipy neutral lipids staining, CD36-postive, nuclear Draq5-negative⁶, and with typically polarized light scattering). Shown are results of 1 of 2 independent experiments.