Fig 2: Dpp4+ progenitor cells contribute to basal adipogenesis in subcutaneous depots. (A) Flow cytometric analysis of SVF cells (CD45(-)/DAPI(-)) from subcutaneous WAT of Dpp4CreER; mGFP mice at the pulse timepoint. mGFP+ lineage traced cells are shown as a green overlay on top of mGFP(-) cells, which are depicted in gray. (B) IHC staining of subcutaneous inguinal WAT (iWAT) at the pulse timepoint. [mGFP: lineage traced cells (green), cells currently expressing DPP4 (magenta), PLIN1: mature adipocytes (red), DAPI: nuclei (blue); scale bar = 380 µM]. Individual antibody staining is represented in grayscale on the right-side inset panels (inset scale bar = 100 µM). (C) IHC staining of Dpp4CreER iWAT (top panels) and axillary WAT (axWAT, bottom panels) following 18 weeks of feeding chow (left) or HFD.

Fig 3: Dpp4+ progenitors contribute extensively to adipogenesis in the murine omentum. (A) Gross histology showing the anatomic location of the mouse omentum. The murine greater omentum is a translucent sheet of collagen-filled connective tissue (reticular interstitium) that extends from the greater curve of the stomach and is draped over the pancreas and intestine. The inferior edge of the omentum is filled with adipose tissue (omental adipose). (B) IHC staining of omental adipose tissue from Dpp4CreER; mGFP mice at the pulse timepoint [mGFP: lineage-traced cells (green), DPP4: cells currently expressing DPP4 (magenta), PLIN1: mature adipocytes (red), DAPI: nuclei (blue); scale bar = 380 µM]. Individual antibody staining is represented in grayscale on the right-side inset panels (inset scale bar = 100 µM). (C) H&E and IHC staining of omental WAT from Dpp4CreER; mGFP mice under chow (left panels) or HFD (right panels) feeding conditions.

Fig 4: Dpp4+ progenitors contribute extensively to adipogenesis in retroperitoneal WAT. (A) Gross histology showing the anatomic location of the mouse retroperitoneal WAT. Murine retroperitoneal WAT is contained within a sheath of reticular interstitial connective tissue that affixes the depot within the retroperitoneal space against the psoas muscles. (B) IHC staining of retroperitoneal WAT from Dpp4CreER; mGFP mice at the pulse timepoint [mGFP: lineage traced cells (green), DPP4: cells currently expressing DPP4 (magenta), PLIN1: mature adipocytes (red), DAPI: nuclei (blue); scale bar = 380 µM]. Individual antibody staining is represented in grayscale on the right-side inset panels (inset scale bar = 100 µM). (C) H&E and IHC staining of Dpp4CreER retroperitoneal adipose under chow (left panels) or high fat diet (right panels) feeding conditions.

Fig 5: Bola3 knockdown did not affect the lipogenesis process in differentiated beige adipocytes. (A, B) Relative Bola3 mRNA (n=4) and protein levels in beige adipocytes differentiated from inguinal WAT (iWAT) stromal vascular fraction (SVF) cells with infection of mouse Bola3 lentiviral shRNA. (C–E) TG concentration (n=4) and representative images of cell morphology and Oil Red O staining in beige adipocytes with Bola3 knockdown. (F) Protein levels of the lipogenic genes (PPAR?, C/EBPa and Perilipin 1) in beige adipocytes infected with mouse lentiviral Bola3 shRNA. (G) Quantitative PCR analysis of the lipogenic genes (Fabp4, Cidec, Glut4, Adipoq, Pparg and Plin1) expression in beige adipocytes infected with lentivirus expressing shBola3 or control (n=4). Data were presented as means ± SEM. *p < 0.05 and ***p <0.001. The results are representative of at least three independent repeats in the cell experiments.