Extended Data Fig. 2: Discrepancies of the two studies4,9 regarding the sorting and culture of Aregs.
From: Identification of a regulatory pathway inhibiting adipogenesis via RSPO2
a) Representative flow cytometry dots plots showing the gating strategy used to identify CD142-, CD142 + , and CD142 + + cells. b) Relative mRNA levels of P3 specific marker genes (Cd142, Gdf10, Igfbp3, Clec11a) in different cell populations. n = 4 biological replicates. Data show the mean ± SEM. c) Histogram plots shows expression level of CD142, CD55, and VAP1 in CD142- cells, CD142 + cells and CD142 + + cells. d) Relative mRNA levels of P1 specific marker genes (Cd55, Pcsk6, Efhd1, Pi16, Smpd3) in different cell populations. n = 4 biological replicates. Data shown as mean ± SEM. e) Relative mRNA levels of P2 specific marker genes (Vap1, Col4a1, Sparcl1, Col18a1, Sdc1) in different cell populations. n = 4 biological replicates. Data shown as mean ± SEM. f) Cells were sorted from ingWAT. Adipogenesis was induced by A Cocktail (1 μM dexamethasone, 0.5 mM isobutylmethylxanthine, 1 μM insulin) or B Cocktail (1 μM dexamethasone, 0.5 mM isobutylmethylxanthine, 125 nM Indomethacin, 1 nM T3, and 20 nM insulin). Quantification of adipogenesis (top) and cell numbers (below) in culture well on differentiation day 6. Data shown as mean ± SEM. F(3,30)= 14.78, P < 0.0001 by two-way ANOVA. For Cocktail difference, F(1,10)=290.5, P < 0.0001. For cell type difference, F(1.929,19.29)=148.8, P < 0.0001. g) Microscopy images of different cell populations shown on differentiation day 6 in f. In all panels, nuclei were stained with Hoechst 33342 (blue) and lipids were stained with LD540 (yellow). Scale bars, 100μm. This figure is related to Fig. 2.
