rabbit anti gamma h2ax antibody  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: RNA polymerase loss by nuclear rupture drives LMNA cardiomyopathy

doi: 10.64898/2026.04.03.716433

Figure Lengend Snippet: A) Immunofluorescence for gamma-H2AX in cardiomyocytes expressing NLS-tdTomato and GFP-icGAS isolated from mice at day 14 post tamoxifen. Texts and icons below images indicate nuclear states. Scale bar: 5 μm. B) Nuclear gamma-H2AX intensity by nuclear states. Density and box plots (interquartile range): signal distribution of all affiliated nuclei. Circles: mean intensity within individual biological replicates (color coded). Statistics: P < 0.05 (*) or P≥0.05 (N.S.) from t-tests on linear regression-estimated means with mouse-clustered standard errors. Underlying data: 164 intact nuclei from 3 WT mice, 152 intact, 116 ruptured, 50 resealed nuclei from 3 Lmna CKO mice. C) Relationship between gamma-H2AX intensity and NLS-tdTomato intensity in nuclei of Lmna CKO cardiomyocytes (340 nuclei from 3 mice). Line: simple linear regression fit with 95% confidence interval. R: Pearson’s correlation coefficient. P: t-test p -value on linear regression-estimated means with mouse-clustered standard errors.

Article Snippet: Rabbit anti-gamma-H2AX antibody (Cell signaling, 9718); mouse anti-BrU/BrdU antibody (BD Biosciences, 555627); rabbit anti-RNA Pol II N-terminal domain (NTD) antibody (Cell signaling, 14958); rat anti-RNA Pol II CTD phospho-Ser5 antibody (Chromotek, AB_2631404); rat anti-RNA Pol II CTD phospho-Ser2 antibody (Millipore Sigma, 04-1571); mouse anti-Lamin A/C antibody (Santa Cruz, sc-376248); rabbit anti-Lamin A antibody (Abcam, ab26300); rabbit anti-PCM1 antibody (Sigma, HPA023370); mouse anti-BANF1 antibody (Abnova, H00008815-M01); rabbit anti-LEMD2 antibody (Millipore Sigma, HPA017340); rabbit anti-CHMP4B antibody (Proteintech, 13683-1-AP); rabbit anti-CHMP7 antibody (Proteintech, 16424-1-AP); mouse anti-VPS4 antibody (Santa Cruz, sc-133122); and mouse anti-c-Myc antibody (Invitrogen, 13-2500).

Techniques: Immunofluorescence, Expressing, Isolation

Journal: EMBO Reports

Article Title: Inhibition of the minor spliceosome restricts the growth of a broad spectrum of cancers

doi: 10.1038/s44319-025-00511-8

Figure Lengend Snippet: ( A ) RT-PCR analysis of MIG splicing changes in A549 cells after 72 h treatment with 10 nM ASO. Schematic depictions of the obtained amplicons are shown on the right with the minor intron in red and the upstream and downstream exons coloured blue and orange, respectively. Exons not separated by a minor intron are grey. ( B ) Representative images of γH2Ax (Ser139) staining in A549 cells 72 h after 10 nM ASO treatment. Scale bar is 10 µm.

Article Snippet: IF & FACS: Rabbit monoclonal anti gamma-H2AX (Ser139) (20E3) , Cell Signaling Technologies , Cat#9718; RRID: AB_2118009.

Techniques: Reverse Transcription Polymerase Chain Reaction, Staining

Journal: Advanced Functional Materials

Article Title: Alleviation of Aging‐Related Hallmarks in a Mouse Model of Progeria via a Nanoparticle‐Based Artificial Transcription Factor

doi: 10.1002/adfm.202425944

Figure Lengend Snippet: Figure 4. Amelioration of aging-associated hallmarks in LMNA G608G cells via Oct4-nanoscript. A) Immunofluorescence for Oct4+ and H3K9me3+ cells in control and LMNAG/G fibroblasts with or without Oct4-nanoscript. Scale bar = 20 μm. B) Quantification of the fluorescence intensity of H3K9me3 in nuclei using a single confocal section. Data represent mean ± SEM. Two-way ANOVA-test, *P < 0.05, **P < 0.01; n = 12 from three samples per group. C) The relative intensity of H3K9me3 levels in control and LMNAG/G fibroblasts treated with Oct4-nanoscript. Data represent mean ± SEM. Two-way ANOVA-test, **P < 0.01; n = 3 per group. D,E) Western blot analysis of 𝛾-H2AX and total histone H3 in control and LMNAG/G fibroblasts treated with

Article Snippet: The cells were incubated overnight at 4 °C with following primary antibodies: anti-Oct4 (Abcam, ab18976; Santacruz, sc-5279), anti-H3K9me3 (Abcam, ab8898), anti-H4K20me3 (Abcam, ab78517), anti-phospho-gamma H2AX (Cell signaling, 9718), anti-Lamin B (Santacruz, sc-374015), antiLamin A/C (BD Biosciences, 612 162), anti-p16INK4A (Invitrogen, PA520379), and anti-α-SMA (Invitrogen, 14-9760-82).

Techniques: Control, Western Blot

Journal: Advanced Science

Article Title: Enhancer Extrachromosomal Circular DNA ANKRD28 Elicits Drug Resistance via POU2F2‐Mediated Transcriptional Network in Multiple Myeloma

doi: 10.1002/advs.202415695

Figure Lengend Snippet: EccANKRD28 dynamics are correlated with the development of VRd resistance and amplify transcription activity. A) CRISPR/Cas9‐mediated generation of custom eccDNA in RPMI‐8226 cells. Abbreviations: CRISPR/Cas9, clustered regularly interspaced short palindromic repeats; NHEJ, non‐homologous end joining. The graphical scheme was created with BioRender. B) Left: The representative colony formation images of RPMI‐8226, U266, and OPM‐2 cells between WT and BR groups. Right: Colony formation rates were expressed as the percentage of colonies in BR cultures compared with that in WT cultures. Each assay was performed in triplicate. C) Representative dual immunofluorescence‐FISH images for eccANKRD28 (green) and γH2AX (red) protein expression in U266 wild‐type (WT) and bortezomib‐resistant (BR) cells. Scale bar, 10 µm. D) Dose‒response curves for the RPMI‐8226 wild‐type (WT), RPMI‐8226 bortezomib‐ or lenalidomide‐resistant (BR or LR) and RPMI‐8226 eccDNA‐CRISPR/Cas9 (EC) cell lines used to determine sensitivity to bortezomib and lenalidomide. The cells were incubated with bortezomib and lenalidomide at serial dilutions, starting at doses of 5 n m bortezomib and 5 µ m lenalidomide, for 48 h before cell viability was determined via a CCK‐8 assay. Results are shown as the mean ± SD of three independent experiments. Statistical significance was assessed using two‐way ANOVA ( * P < 0.05; ** P < 0.01; *** P < 0.001). E) Multiple correlation analysis between the IC 50 and eccANKRD28 copy number in RPMI‐8226, U266, and OPM‐2 cells. F) Mean eccANKRD28 copy number after eccANKRD28 induction on day 0 ± BTZ treatment beginning on day 3. G) Interphase FISH microscopy for eccANKRD28. Case #1 and Case #2 were representative images of CD138+ sorted plasma cells from NDMM, Case #1 was evaluated as MRD‐neg (HS group, n = 5), and Case #2 was evaluated as SD (LS group, n = 5) after three cycles of the VRd regimen. Representative images from a series of 10 images of each subject. Scale bar, 10 µm. H‐I) Copy number of eccANKRD28 and ANKRD28 mRNA expression determined by qPCR in the HD/HS/LS group. ** P < 0.01; *** P < 0.001.

Article Snippet: Anti‐gamma H2AX (#9718T) was purchased from Cell Signaling Technology (Beverly, USA).

Techniques: Activity Assay, CRISPR, Non-Homologous End Joining, Immunofluorescence, Expressing, Incubation, CCK-8 Assay, Microscopy, Clinical Proteomics