Fig 2: Diagram presentation of the molecular mechanism. KDM3A encourages the transcription of ETS1 through the demethylation and histone modification of H3K9me2, while ETS1 further binds to the promoter region of KIF14 to promote its transcription activity, which activate the Hedgehog signaling pathway and promote CC progression.

Fig 3: Expression of HUWE1 in CD4+ T cells in peripheral blood from immune thrombocytopenic purpura patients. (A) Quantitative real-time PCR (qRT-PCR) and Western blot assays were performed to detect the mRNA and protein levels of HUWE1 in CD4+ T cells in the peripheral blood from healthy controls and immune thrombocytopenic purpura (ITP) patients; and the quantitative analysis of HUWE1 protein level (mean ± SEM, n = 5). (B) Correlation analysis of the mRNA level of HUWE1 and platelet counts in CD4+ T cells in the peripheral blood from ITP patients (r = −0.890, p < 0.01), n = 30. (C) Flow cytometry was applied to analyze the percentage of Treg cells in CD4+ T cells in peripheral blood from ITP patients and correlation analysis of the mRNA level of HUWE1 and the percentage of Treg cells in CD4+ T cells in peripheral blood from ITP patients (r = −0.858, p < 0.01), n = 30. (D) Western blot was performed to assess the Ets-1 protein level in the CD4+ T cells in the peripheral blood from ITP patients. The experiment was repeated three times. GAPDH is applied as the loading control. **p < 0.01 vs. control. ITP, immune thrombocytopenic purpura.

Fig 4: Overexpression of miR-128 blocks the promotion of ETS1 on osteogenic differentiation. (A) Transfection efficacy of miR-128 mimic determined by RT-qPCR (unpaired t-test, *p < 0.05). (B) mRNA and protein levels of osteogenic markers including OCN, OSX and RUNX2 in cells determined by RT-qPCR and western blot analysis, respectively (two-way ANOVA, *p < 0.05). (C) ALP concentration in MC3T3-E1 cells measured by ALP staining (unpaired t-test, *p < 0.05). (D) Calcareous accumulation in MC3T3-E1 cells measured by ARS staining (unpaired t-test, *p < 0.05).

Fig 5: Inhibition of ß-catenin blocks the promotion of oe-ETS1 on osteogenic differentiation. (A) Protein expression of ß-catenin in MC3T3-E1 cells after CWP232228 administration determined by western blot analysis (one-way ANOVA, *p < 0.05). (B) mRNA and protein expression of RUNX2, OCN, and OSX in cells on day 7 determined by RT-qPCR and western blot analysis, respectively (two-way ANOVA, *p < 0.05). (C) ALP concentration in cells measured by ALP staining (unpaired t-test, *p < 0.05). (D) Calcareous accumulation in cells on day 21 evaluated by ARS staining (unpaired t-test, *p < 0.05).