Fig 2: ASD ameliorated oxidative stress via modulation of Nrf2/JunD/p65 signaling in rats with TAA-induced liver injury. (A) Representative Western blot images and (B) histograms showing the effect of TAA or TAA+ASD on the expression of nuclear-translocated Nrf2, p65, and JunD in rats with TAA-induced liver injury. Histograms of the effect of TAA or TAA+ASD on the (C) GSH/GSSG ratio, (D) 4-hydroxynonenal, or (E) MDA levels in the liver injury model rats. PCNA was used as nuclear marker and as loading control. PCNA – proliferating cell nuclear antigen; GSH – glutathione; GSSG – glutathione disulfide; MDA – malondialdehyde. * p<0.05; ** p<0.01; *** p<0.001.

Fig 3: KEGG enrichment of 249 DEGs identified between chemotherapy sensitive and insensitive OC. (A) MAPK signaling pathway including RASGRF1, MAPT, FOS, FOSB, DUSP family, JUN, JUND, JUNB, NR4A1, and GADD45G. (B) ERBB signaling pathway including NRG1, EREG, HBEGF, PAK3, and JUN. (C) IL17 signaling pathway including JUN, JUND, JUNB, FOS, FOSB, S100A7, CXCL2/3, and MUC5B. (D) TNF signaling pathway, including JUN, JUND, JUNB, FOS, FOSB, CXCL2/3, LTA, and SOCS3. Common DEGs in chemotherapy sensitivity group and ovarian cancer occurring group were shown by red letter

Fig 4: Jund maintains appropriate accessibility of cell type–specific chromatin landscapes. (A) UMAP plots of VEC, AEC, HEC, L/M-HSPC, and E-HSPC in integrating scATAC-seq data of WT and jund−/− cells. (B) GO enrichment analysis showing the functional annotation of genes associated with gained accessible peaks in each cell type of jund−/− embryos compared with WT. (C) GO enrichment analysis showing the functional annotation of genes associated with reduced accessible peaks in each cell type of jund−/− embryos compared with WT. (D) Bar plot showing the altered frequency of marker peaks in WT and jund−/− embryos, compared AEC with VEC, and HEC with AEC.

Fig 5: Jund and Hoxa9a jointly inhibit the activity of dll4 enhancer through depleting H3K27ac around binding sites. (A) Bar plot depicting the genomic locations and chromatin states of Jund binding peaks. Chromatin states were classified into five categories: active (high H3K4me3 or H3K27ac with low H3K27me3), poised (high H3K4me3 and H3K27me3), repressed (high H3K27me3 with low H3K4me3 and H3K27ac), mixed, and unmarked. (B) Jund binding sites and H3K27ac peak signals at dll4 genomic locus. The putative enhancer of dll4 is shaded. (C) ChIP-qPCR analysis showing relative enrichment of dll4 enhancer in IgG and anti-Jund groups in HECs sorted from WT and jund−/− embryos with Tg(kdrl:mCherry/runx1:en-GFP) background. (D) ChIP-qPCR analysis showing relative DNA level of dll4 enhancer in H3K27ac and H3K27me3 groups in HECs sorted from WT and jund−/− embryos with Tg(kdrl:mCherry/runx1:en-GFP) background. (E) WISH (Left) and quantification (Right) showing the expression of dll4 in the AGM region of WT and jund−/− embryos at 36 hpf. (F) Luciferase reporter assays of dll4 enhancer construct in WT and jund−/− embryos under different conditions of flag-jund mRNA and HA-hoxa9a mRNA injections. (G) ChIP-qPCR analysis showing relative enrichment of dll4 enhancer in IgG and anti-HA-Hoxa9a groups in HECs sorted from Tg(kdrl:mCherry/runx1:en-GFP) embryos. (H) Western blotting analysis of Flag-Jund and HA-Hoxa9a protein binding to biotin-labeled WT or Mut dll4 enhancer probes. Proteins were purified from HECs/HSPCs sorted from Tg(runx1:en-GFP) embryos injected with flag-jund or HA-hoxa9a mRNA. (I) Western blotting analysis of HA-Hoxa9a protein binding to biotin-labeled WT dll4 enhancer probes. Proteins were purified from HECs/HSPCs sorted from WT or jund−/− embryos in Tg(runx1:en-GFP) background injected with HA-hoxa9a mRNA. (J) qPCR showing the relative mRNA expression of dll4 in HECs sorted from WT and jund−/− embryos in Tg(kdrl:mCherry/runx1:en-GFP) background injected with control MO or hoxa9a MO at 36 hpf. (K) ChIP-qPCR analysis showing relative DNA level of dll4 enhancer in H3K27ac and H3K27me3 groups in HECs sorted from control embryos, hoxa9a crispants, and hoxa9a morphants in Tg(kdrl:mCherry/runx1:en-GFP) background. (L) WISH (Left) and quantification (Right) showing the expression of HEC and HSPC markers (gfi1aa, runx1, and cmyb) in the AGM region of WT embryos injected with control MO, jund−/− embryos injected with control MO and jund−/− embryos injected with dll4 MO at 36 hpf. (M) Confocal imaging (Left) and statistical data (Right) of Tg(kdrl:mCherry/runx1:en-GFP) showing the number of kdrl+runx1+ HECs (white arrowheads) in the AGM region of WT embryos injected with control MO, jund−/− embryos injected with control MO and jund−/− embryos injected with dll4 MO at 36 hpf. (N) qPCR showing the relative mRNA expression of runx1 and gfi1aa in HECs, and runx1, cmyb, gata2b, and gfi1b in HSPCs sorted from WT or jund−/− embryos in Tg(kdrl:mCherry/runx1:en-GFP) background injected with control MO or dll4 MO at 36 hpf. Error bar, mean ± SD. n ≥ 3 biological replicates. Two-tailed unpaired Student’s t test, ns: no significance, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (C–G and J–N). Arrowheads denote marker gene-positive signals (E and L). Numerical values represent embryos displaying the characteristic phenotype (numerator) relative to total embryos analyzed (denominator) for each group (E and L). [Scale bars, 100 μm (E and L), 50 μm (M).]