Fig. 5: Rspo2 inhibits transition of eP1 cells to eP2 cells.

a–f, Experimental scheme (a) for transplantation of tdTomato⁺ eP1 cells into inguinal adipose tissue of wild-type (WT) mice. FACS analysis (b) of VAP1 and CD142 expression in tdTomato⁺ eP1 cells 10 d after transplantation. Expression of P1 marker genes (c) (Cd55, Dpp4, Pi16 and Psck6), P2 marker genes (d) (Vap1, Icam1, Col4a1 and Sparcl1), Pparg and Cebpa (e) and P3 marker genes (f) (Cd142, Gdf10, Clec11a and Igfbp3) in eP1 cells (from donor mice), eP2 cells (from donor mice), eP3 cells (from donor mice), VAP1⁺ cells (derived from implanted eP1 cells) and VAP1⁻ cells (derived from implanted eP1 cells). Data are shown as mean ± s.e.m., n = 4 biological replicates. g–m, Experimental scheme (g) for injection of AAVs into ingWAT for overexpression of RSPO2. Western blot images (h) and quantification (i) of RSPO2 protein and Rspo2 mRNA (j) in ingWAT. FACS analysis of eP1/SVF (k), eP2/SVF (l), CD55⁺VAP1⁺ (m) in ingWAT. Data are shown as mean ± s.e.m., n = 6 mice (h,i), n = 5–6 mice (j), n = 5 mice (k–m). Data were analyzed using two-tailed Student’s t-test. n,o, Experimental scheme (n) for transplantation of tdTomato⁺ eP1 cells into RSPO2 overexpression mice. FACS analysis of (VAP1⁺:tdTomato⁺) cells in tdTomato⁺ eP1 cells (o). Data are shown as mean ± s.e.m., n = 5 biological replicates. Data were analyzed using a two-tailed paired Student’s t-test.

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