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. 2023 Aug 23;12:RP85380. doi: 10.7554/eLife.85380

Figure 7. FOXC2 is essential for sustaining the quiescent state of spermatogonial stem cells (SSCs) via regulating cell cycle.

Figure 7.

(A) Workflow schematic illustration of the CUT&Tag_FOXC2 analysis on the fluorescence-activated cell sorting (FACS)-sorted FOXC2+ cells. (B) Pie chart for CUT&Tag_FOXC2 peaks genome distribution. (C) Profiling of CUT&Tag_FOXC2 peaks in proximity to transcriptional starting site (TSS). The distance to TSS within 1000 was highlighted in the purple box. (D) Top Gene Ontology (GO) terms enrichment by genes annotated by CUT&Tag_FOXC2 peaks. (E) Venn diagram of FOXC2 target genes defined by overlapping the CUT&Tag sequencing and single-cell RNA-sequencing (scRNA-seq) datasets. (F) GO terms enrichment by the FOXC2 target genes related to cell cycle regulation. (G) Chromatin landscapes of CUT&Tag_FOXC2 peaks of the candidates associated with negative cell cycle regulation. (H) The DNA-binding motif for FOXC2 (predicted with HOMER). (I) The cell cycle-related candidates possessing high binding potential (>0.8, predicted with JASPAR SCAN). (J) CUT&Tag-qPCR validation of the cell cycle arrest regulatory genes. (n=3). Values, mean ± s.e.m.; p-values were obtained using two-tailed t-tests (****p-value <0.0001). (K) The model for the maintenance of the FOXC2+ SSC subpopulation in adult testis.

Figure 7—source data 1. Excel spreadsheet with the list of the differentially expressed genes found by Cleavage Under Targets and Tagmentation (CUT&Tag) sequencing and Gene Ontology terms of the 306 crossed candidates.