Fig 1: Upregulation of H3K27me3 demethylase KDM6B is detected in MCAO-operated mice and OGD/R-exposed astrocytes(A) The KDM6B mRNA expression in astrocytes after OGD/R treatment determined by qRT-PCR. (B) The KDM6B protein expression in OGD/R-exposed astrocytes measured by western blot analysis. (C) The KDM6B mRNA expression in mouse brain tissues 1 day post-MCAO determined by qRT-PCR. (D) The KDM6B protein expression in mouse brain tissues 1 day post-MCAO measured by western blot analysis. Data were measurement data and expressed by mean ± standard deviation. Data between two groups were analyzed by unpaired t test, and the experiment was repeated three times. **p < 0.05.
Fig 2: Effects of GSK-J4 or DZNeP on prostate cancer cell lines(A) Cell line viability treated with GSK-J4. PC-3, LNCaP and DU 145 cells were treated for 24 h, 48 h and 72 h with increasing concentrations of GSK-J4. Cell viability was assessed using the XTT assay. The percentage of viable cells was determined as percent of viability of untreated cells. Values shown are the average (mean ± S.E.M) from quadruplicate samples for each incubation condition. (B–C) Western-blot analysis of GSK-J4 and DZNeP efficacity. Cells were treated at IC50 concentration (PC-3: 3.53 µM, LNCaP: 3.93 µM and DU 145: 22.87 µM for 48 h for GSK-J4 and 10 µM for 72 h for DZNeP) (TT) or untreated (N). Quantification representation were expressed as relative fold change in protein expression of JMJD3 and EZH2 in response to GSK-J4 or DZNeP exposure respectively after normalization to GAPDH density.
Fig 3: Epigenetic regulation of HOXA5 occurs at its putative promoter region. (A) Schematic depiction of the HOXA5 locus on human chromosome 7. Boxes represent exons, lines represent introns, and arrows show the direction of transcription. The gray bars represent the amplicon sites used in chromatin immunoprecipitation-PCR (ChIP-PCR). (B) ChIP-PCR analysis of the histone modifications H3K4me3, H3K27me3, and H3K9ac in MCF7 and TAMR cells. (C) Western blotting images showing protein levels of the epigenetic modifiers in MCF7 and TAMR cells. ß-Actin was used as an internal control. (D) ChIP-PCR analysis of EZH2, SUZ12, EED, JMJD3, and UTX in MCF7 and TAMR cells. All experiments were performed in triplicate. *** p < 0.001 compared with MCF7 by Student's t-test.
Fig 4: The mechanism of JMJD3.The mechanism of JMJD3 in the tumor cell arrest.
Fig 5: Interaction of GSK-J4 and DZNeP on PTEN and AR pathwaysEffects of JMJD3 and EZH2 inhibitors on key pathways involved in prostate cancer. JMJD3 inhibitor GSK-J4 enhanced H3K27me3 which inhibited PTEN expression and activated AKT; by contrast, DZNeP counteracted these effects. GSK-J4 acts on AR-driven transcription and interferes with proliferation. Arrows indicate an activation, blocked arrows indicate an inhibition and dotted arrows a presumed interaction.
Supplier Page from Abcam for Anti-KDM6B / JMJD3 antibody