primary antibody phosphorylation of histone h3 at serine 10 (p-h3(s10)) monoclonal antibody 53348  (Cell Signaling Technology Inc)

Journal: Molecules

Article Title: Colchicine Binding Site Tubulin Inhibitors Impair Vincristine-Resistant Neuroblastoma Cell Function

doi: 10.3390/molecules30102186

Figure Lengend Snippet: Induction of mitotic and apoptotic protein markers across neuroblastoma cell lines. ( a ) Western blots of protein lysates collected 24 h after treatment of Kelly cells with 0.02% DMSO, 100 nM colchicine, 100 nM 4h, and 500 nM 4k. GAPDH is used as a loading control. P-S10 H3 and CYCLIN B1 are mitotic markers, and cleaved PARP is an apoptotic marker. ( b ) Western blots of protein lysates prepared as in ( a ) from SK-N-AS and SK-N-SH cell lines. Uncropped Western blots used in this figure are included within .

Article Snippet: The antibodies used were as follows: c-MYC (Abcam, Cambridge, UK, AB32072), N-MYC (Cell Signaling, Waltham, MA, USA, 51705), GAPDH-HRP (Cell Signaling, 8884), p-S10-H3 (Cell Signaling, 53348), β-TUBULIN (Cell Signaling, 2128), β3-TUBULIN (Cell Signaling, 5568), cleaved PARP (Cell Signaling, 9541), CYCLIN B1 (Cell Signaling, 4138), ABCB1 (Cell Signaling, 13342), and ABCC1 (Cell Signaling, 72202).

Techniques: Western Blot, Control, Marker

Journal: bioRxiv

Article Title: Potent synthetic lethality between PLK1 and EYA-family inhibitors in tumours of the central and peripheral nervous system

doi: 10.1101/2025.01.19.633804

Figure Lengend Snippet: (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, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (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, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (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 outline. (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 foci number in S-phase following treatment with DMSO, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (one-way ANOVA with Tukey’s multiple comparison test, ** p ≤ 0.01). (H) Quantitation of RAD51 foci intensity in S-phase following treatment with DMSO, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (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, Protein Tech 22658 (1:1000), EYA2, Millipore HPA027024 (1:1000), EYA3, Protein Tech 21196 (1:1000), EYA4, Santa Cruz SC393111 (1:1000), Actin, Sigma a2066 (1:2000), PARP, Cell Signalling 9542 (1:1000), gamma-H2AX Millipore 05-636 (1:1000), Vinculin, Sigma V9131 (1:4000), p-S10 H3 Cell Signalling 9706 (1:1000), CD133, Cell Signalling 64326 (1:1000), and Sox9, Cell Signalling 82630 (1:1000).

Techniques: Immunofluorescence, Staining, Cytometry, Quantitation Assay, Comparison

Journal: bioRxiv

Article Title: Potent synthetic lethality between PLK1 and EYA-family inhibitors in tumours of the central and peripheral nervous system

doi: 10.1101/2025.01.19.633804

Figure Lengend Snippet: (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, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (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, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (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 outline. (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 foci number in S-phase following treatment with DMSO, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (one-way ANOVA with Tukey’s multiple comparison test, ** p ≤ 0.01). (H) Quantitation of RAD51 foci intensity in S-phase following treatment with DMSO, Benzarone (7.5 µM), Bi2536 (10 nM), or Benzarone + Bi2536 for 5.5 hrs (one-way ANOVA with Tukey’s multiple comparison test, * p ≤ 0.05, ** p ≤ 0.01, **** p ≤ 0.0001).

Article Snippet: Cells were incubated with anti-p-S46 TCTP (Cell Signalling 5251) and anti-p-S10 H3 primary antibodies (Cell Signalling 9701) overnight at 4 ° C followed by 3 x 10 minute washes in PBS.

Techniques: Immunofluorescence, Staining, Cytometry, Quantitation Assay, Comparison