anti p s10 h3  (Cell Signaling Technology Inc)

Journal: Genes & Development

Article Title: Potent synthetic lethality between PLK1 and EYA family inhibitors in tumors of the central and peripheral nervous systems

doi: 10.1101/gad.353271.125

Figure Lengend Snippet: Combined EYA and PLK1 inhibition additively decreases PLK1 substrate phosphorylation in G2 and M phase and RAD51 focus formation and intensity in S phase. ( A ) Representative immunofluorescence images showing G2- and M-phase cells stained with p-S46 TCTP, p-S10 H3, and DAPI. ( B ) Representative quantitative image-based cytometry plot showing G2- and M-phase cell cycle gates as determined by staining intensity of p-S10 H3 and DAPI. ( C ) Cellular quantitation of p-S46 TCTP intensity in G2 phase following treatment with DMSO, 7.5 µM benzarone, 10 nM Bi2536, or benzarone + Bi2536 for 5.5 h. One-way ANOVA with Tukey's multiple comparison test. (**) P ≤ 0.01, (****) P ≤ 0.0001. ( D ) Cellular quantitation of p-S46 TCTP intensity in M phase following treatment with DMSO, 7.5 µM benzarone, 10 nM Bi2536, or benzarone + Bi2536 for 5.5 h. One-way ANOVA with Tukey's multiple comparison test. (*) P ≤ 0.05, (**) P ≤ 0.01, (****) P ≤ 0.0001. ( E ) Representative immunofluorescence images showing an S-phase cell stained with RAD51, EdU, and DAPI. Individual RAD51 foci are indicated with red outlines. ( F ) Representative quantitative image-based cytometry plot showing S-phase cell cycle gate as determined by staining intensity of EdU and DAPI. ( G ) Cellular quantitation of RAD51 focus number in S phase following treatment with DMSO, 7.5 µM benzarone, 10 nM Bi2536, or benzarone + Bi2536 for 5.5 h. One-way ANOVA with Tukey's multiple comparison test. (**) P ≤ 0.01. ( H ) Quantitation of RAD51 focus intensity in S phase following treatment with DMSO, 7.5 µM benzarone, 10 nM Bi2536, or benzarone + Bi2536 for 5.5 h. One-way ANOVA with Tukey's multiple comparison test. (*) P ≤ 0.05, (**) P ≤ 0.01, (****) P ≤ 0.0001.

Article Snippet: Western blot primary antibodies targeted EYA1 (1:1000; Protein Tech 22658), EYA2 (1:1000; Millipore HPA027024), EYA3 (1:1000; Protein Tech 21196 ), EYA4 (1:1000; Santa Cruz Biotechnology SC393111), Actin (1:2000; Sigma a2066), PARP (1:1000; Cell Signaling Technology 9542), γ-H2AX (1:1000; Millipore 05-636), vinculin (1:4000; Sigma V9131), p-S10 H3 (1:1000; Cell Signaling Technology 9706), CD133 (1:1000; Cell Signaling Technology 64326), and Sox9 (1:1000; Cell Signaling Technology 82630).

Techniques: Inhibition, Phospho-proteomics, Immunofluorescence, Staining, Cytometry, Quantitation Assay, Comparison

Journal: bioRxiv

Article Title: Live dynamics of induced cell-cell fusion between mitotic and interphasic cells

doi: 10.64898/2026.01.27.700572

Figure Lengend Snippet: (A) Comparison of Mad2 localization in control prometaphase and anaphase cells and mitotic fused cells undergoing premature mitotic exit. Mad2 can be detected at some kinetochores despite the presence of nuclear envelope membranes. (B) Time lapse of cerulean-Cyclin B1 transiently transfected in LLC-PK1 cells expressing H2B-mCherry after cell-cell fusion. Cyclin B1 was not degraded during induced mitotic exit of the mitotic cell. (C) Analysis of pH3-s10 in fixed prometaphase control cells and at different time points after cell-cell fusion. At 30 minutes after cell-cell fusion there is a mixed population of cells with high (second panel) and low (third panel) pH3-s10 levels, despite the presence of nuclear envelope membranes. At 60 minutes after cell-cell fusion, most cells show very low levels of pH3-s10. Magenta squares show magnification of nuclear envelope membranes around chromosomes. (D) Quantification of pH3-s10 in telophase control cells and at several time points after cell-cell fusion. Control, n=19 cells; 20 min, n=20 cells; 30 min, n=22 cells; 40 min, n=27 cells; 50 min, n=32 cells and 60 min, n=26 cells. Note that only 50 and 60 minutes after cell-cell fusion the levels of pH3-s10 become similar to control levels. Statistics, non-parametric Mann-Whitney test. Time is h:min. Scale bar is 10μm.

Article Snippet: Primary mouse mab414 (1:1000; ab24609, Abcam), mouse anti-α-Tubulin (1:500, DM1A, sigma), mouse anti-Mad2 (1:100, sc-65492, Santa Cruz) and rabbit anti-pH3-s10 (1:200, 3377, Cell Signaling).

Techniques: Comparison, Control, Transfection, Expressing, MANN-WHITNEY