Fig. 3: RNA sequencing analyses of GCs isolated from the ovaries of mice treated with or without CTX for 6 h.

A Volcano plot comparing the transcripts of the GCs isolated from the ovaries of mice treated with or without CTX for 6 h. Transcripts that increased or decreased by more than 2-fold in the GCs isolated from the ovaries of mice treated with CTX were highlighted in red or blue, respectively. GCs isolated from 3 mice were pooled. N = 3 biological replicates. B Heatmap of differentially expressed genes in the GCs isolated from the ovaries of mice treated with or without CTX for 6 h sorted by adjusted P value by plotting their log₂ transformed expression values in replicates. C, D GO (C) and GSEA (D) analysis of significantly enriched pathways in the upregulated gene sets of GCs isolated from the ovaries of mice treated with CTX. E Relative expression of cyclin-dependent kinase inhibitor 1A (Cdkn1a, also known as p21), ectodysplasin A2 receptor (Eda2r), TNF receptor superfamily member 6/Fas cell surface death receptor (Fas), cyclin G1 (Ccng1), growth arrest and DNA-damage-inducible 45 gamma (Gadd45g), arachidonate 5-lipoxygenase (Alox5), early growth response 4 (Egr4), Kruppel-like transcription factor 4 (Klf4), and BTG anti-proliferation factor 2 (Btg2) mRNA levels by RT-qPCR in the GCs isolated from the ovaries of mice treated with or without CTX. N = 6 mice per time point for each group. Data are the mean ± SD of at least three independent experiments. Statistical analyses were carried out using two-tailed Student’s t-test; n.s. non-significant; ***P < 0.001.