Prkar2b and Marcks were two representative markers for indicating the imbalance of GCs composition in obese mice
(A) qPCR assay of GCs isolated from RD or HFD mouse growing follicles for representative up-regulated genes identified in Figure 2A. Expression levels were normalized to Rps24. Two technical duplicates were performed in individual qPCR experiments, and the representative result of three independent biological replicates is shown. Error bars indicate the mean ± SEM.
(B) Immunofluorescence staining for Prkar2b and Marcks in ovarian tissues from RD and HFD mice. Tissues were counterstained with DAPI and Gdf9 or Ddx4.
(C) Boxplots showing expression levels of Cyp17a1 for ETCs, and Ar for GCs, in RD and the obese mice. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001, n.s. denotes not significant (Wilcoxon rank-sum test).
(D) Trend lines showing relative expression levels for Ar in two branches, State 1–2 and State 1–3, identified by using Monocle2.
(E) Boxplots showing expression levels of Ar for Ldhb+ GCs, Htra1+ GCs, and Inhbb+ GCs, in RD and the obese mice. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001, n.s. denotes not significant (Wilcoxon rank-sum test).