Journal: Journal of Biological Chemistry

Article Title: Identification and characterization of second-generation EZH2 inhibitors with extended residence times and improved biological activity

doi: 10.1016/j.jbc.2021.100349

Figure Lengend Snippet: Figure 3. Exploration of allosteric modulation of EZH2i potencies. A, effect of allosteric modulation on inhibitor residence times for EZH2 inhibitors used to develop kinetic methodologies. Error bars represent propagated error from measured values used in calculation. B, modulation of CPI-1205 residence time by MAK683 (EEDi). Representative data are plotted as the average of two technical replicates (±) standard deviation. The residence time of CPI-1205 for basal PRC2 is 1.2 ± 0.3 h (mean ± SD of nine biological replicates each in duplicate). For basal PRC2 in the presence of 10 μM MAK683, the residence time of CPI-1205 is 0.97 ± 0.1 h (average ±SD of three biological replicates each in duplicate). C, MAK683 (10 μM) inhibition of CPI-1205 residence time enhancement by H3K27me3 activator peptide (10 μM). Representative data are plotted as the average of two technical replicates ± standard deviation. The residence time of CPI-1205 PRC2 in the presence of H3K27me3 activator peptide and MAK-683 (10 mM each) is 1.3 ± 0.2 h (mean ± SD of four biological replicates each in duplicate). D, comparison of association rate constant values of EZH2i (±) 10 μM H3K27me3 activator peptide. Data are individual replicates with error bars representing the standard deviation of the mean. E and F, plots of residence time values for basal (E; R2 0.78) and activated (F; R2

Article Snippet: Twenty-five microliters of αH3K27me3 (0.25 μg/ml, Cell Signaling 9733), which had previously been validated in-house (48) or anti-Histone H3 (0.125 μg/ml, Cell Signaling 4499) in 1% BSA in TBST, was added for 30 min at RT with shaking.

Techniques: Standard Deviation, Inhibition, Comparison

Journal: Journal of Biological Chemistry

Article Title: Identification and characterization of second-generation EZH2 inhibitors with extended residence times and improved biological activity

doi: 10.1016/j.jbc.2021.100349

Figure Lengend Snippet: Figure 7. Biological characterization of second-generation EZH2i. A, plot of inhibitor residence time values versus EC50 values for H3K27me3 reduction in HeLa cells. B, plot of inhibitor residence time values versus GI50 values for KARPAS-422 cell line. C, plot of inhibitor pKi values versus GI50 values in KARPAS- 422 cell line. D, modulation of global H3K27me3 levels (Day 12) in a Karpas-422 xenograft model by CPI-1328 and EPZ-6438. Data are individual replicates with error bars representing the standard deviation of the mean. E and F, efficacy of CPI-1328 and EPZ-6438 in a Karpas-422 xenograft model, respectively. Data are the average of three or more animals with error bars representing the standard error of the mean.

Article Snippet: Twenty-five microliters of αH3K27me3 (0.25 μg/ml, Cell Signaling 9733), which had previously been validated in-house (48) or anti-Histone H3 (0.125 μg/ml, Cell Signaling 4499) in 1% BSA in TBST, was added for 30 min at RT with shaking.

Techniques: Standard Deviation

Journal: International journal of oral science

Article Title: NUP62 alleviates senescence and promotes the stemness of human dental pulp stem cells via NSD2-dependent epigenetic reprogramming.

doi: 10.1038/s41368-025-00362-y

Figure Lengend Snippet: Fig. 5 NUP62 mediates epigenetic reprogramming by upregulating the global levels of H3K36me2 and H3K36me3. a, b Western blot analyses of H3K36me2, H3K36me3, H3K27ac, H3K4me3, and H3K9me3 in human dental pulp stem cells (HDPSCs) transfected with lentiviral vectors or overexpressing NUP62. c, d Immunofluorescence staining and quantification of H3K36me2, respectively (n = 5). Scale bar, 50 μm. e, f Immunofluorescence staining and quantification of H3K36me3, respectively (n = 5). Scale bar, 50 μm. g Heatmap of global H3K36me2 and H3K36me3 CUT&Tag-seq signals from HDPSCs transfected with empty lentiviral vector or NUP62 overexpression lentivirus (n = 2). h Gene track view of normalized bigWig reads at the promoters of SIRT6, HMGA1 and HMGA2 in HDPSCs transfected with a lentiviral vector or overexpressing (n = 2). *P < 0.05, **P < 0.01 and ***P < 0.001

Article Snippet: After blocking in 5% BSA for 30 min, the cells were incubated overnight at 4 °C with primary antibodies against NUP62 (Abcam, ab96134, 1:200), γH2AX (Cell Signaling Technology, #9718T, 1:200), H3K36me2 (Active Motif, 39056, 1:200), H3K36me3 (Cell Signaling Technology, #4909S, 1:200) or E2F1 (Cohesion, CQA8351, 1:100).

Techniques: Western Blot, Transfection, Staining, Plasmid Preparation, Over Expression