Fig 2: The effects between circ_0003074 and miR-516b-5p on the proliferation, apoptosis, migration and invasion of Rg3-treated MG63 cells and Saos2 cells. A–G Rg3-treated MG63 cells and Saos2 cells were transfected with pCD-ciR, circ_0003074, circ_0003074 + miR-NC or circ_0003074 + miR-516b-5p, and cell viability was investigated by CCK-8 (A), cell proliferation by EdU assay (B), cell apoptosis by flow cytometry analysis (C), cell migration by wound-healing assay (D), cell invasion by transwell invasion assay (E), and the protein expression of PCNA, cleaved caspase-3 and MMP9 by Western blotting (F, G). *P < 0.05, **P < 0.01 and ***P < 0.001

Fig 3: MiR-516b-5p regulated Rg3-treated OS cell malignancy through KPNA4. A–G Rg3-treated MG63 cells and Saos2 cells were transfected with anti-miR-NC, anti-miR-516b-5p, anti-miR-516b-5p + si-NC, anti-miR-516b-5p + si-KPNA4, and cell viability was investigated by CCK-8 (A), cell proliferation by EdU assay (B), cell apoptosis by flow cytometry analysis (C), cell migration by wound-healing assay (D), cell invasion by transwell invasion assay (E), and the protein expression of PCNA, cleaved caspase-3 and MMP9 by Western blotting (F, G). *P < 0.05, **P < 0.01 and ***P < 0.001

Fig 4: JNK deficiency promotes early disease lesions.(A) Eighteen weeks after gene deletion, tissue sections were prepared from mammary glands of JNKWT (n = 5) and JNKKO (n = 12) female mice. The mice were not matched for estrus cycle. Representative H and E-stained sections are presented. Scale bar = 100 µm. (B, C) Proliferation was examined in mammary glands at 18 weeks after gene ablation by staining tissue sections with an antibody to PCNA (JNKWT n = 4 mice, JNKKO n = 5 mice). Representative PCNA-stained and DAPI counter-stained glands are presented (B, Scale bar = 50 µm). The percent of PCNA+ cells was quantified (mean ± SEM) (C).10.7554/eLife.36389.031Figure 7—source data 1.Spreadsheet of source data for Figure 7.

Fig 5: The effects between Rg3 and circ_0003074 overexpression on OS cell malignancy. A The efficiency of circ_0003074 overexpression was determined by qRT-PCR. B–I Both MG63 cells and Saos2 cells were treated with 80 µM Rg3, pCD-ciR, circ_0003074, 80 µM Rg3 + pCD-ciR or 80 µM Rg3 + circ_0003074, with untreated cells as a control, and cell viability was investigated by CCK-8 (B), cell proliferation by EdU assay (C), cell apoptosis by flow cytometry analysis (D, E), cell migration by wound-healing assay (F), cell invasion by transwell invasion assay (G), and the protein expression of PCNA, cleaved caspase-3 and MMP9 by Western blotting (H, I). *P < 0.05, **P < 0.01 and ***P < 0.001