phospho p53 ser15 antibody  (Cell Signaling Technology Inc)

Journal: eBioMedicine

Article Title: Modified hTERT treatment ameliorates pressure overload-induced heart failure

doi: 10.1016/j.ebiom.2026.106203

Figure Lengend Snippet: JV101 inhibited telomeric DNA damage, mitochondrial dysfunction and chronic inflammation in TAC-induced HF. (A) Chord diagrams showing differentially expressed genes corresponding to selected regulated pathways in TAC + JV101 versus TAC + Vector comparisons. (B and C) Immunoblotting analysis of multiple inflammatory cytokines in heart of TAC-induced heart failure mice (n = 3 per group). (D) Nuclear lysate isolated from cardiomyocytes perfused from mice of TAC + Vector, TAC + JV101 and Sham group were immunoblotted for gH2AX, Histone H2AX, and 53BP1; Whole cell lysate was immunoblotted for p-p53(Ser15), p53, PGC1 a and TFAM. H3 and GAPDH were used as loading controls. (E–G) Representative transmission electron micrographs of left ventricular myocardium of the indicated study groups. Quantitative analyses of cristae number (F) and mitochondria density (G) in TAC hearts. (H) Quantification of mitochondrial DNA (mtDNA) copy numbers of cardiomyocytes perfused from mice of TAC + Vector, TAC + JV101 and Sham group (n = 6). (I) Intracellular ATP content of TAC + Vector, TAC + JV101 and Sham group (n = 6). Data are represented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Statistical differences among groups were analysed by one-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: The primary antibodies: histone H 2 AX (1:1000, Cell Signalling Technology, 7631S), γH 2 AX (1:1000, Cell Signalling Technology, 9718S), 53BP1(1:1000, Cell Signalling Technology, 4937S), p53 (1:1000, Cell Signalling Technology, 2524S), Phospho-p53 (Ser15) (1:1000, Cell Signalling Technology, 9284s), PGC-1α (1:1000, Santa Cruz, sc-517380), TFAM (1:1000, Proteintech, 22586-1-AP), TRF1 (1:1000, Abcam, ab192629), RAP1 (1:1000, Santa Cruz, sc-53434), TIN2 (1:1000, Proteintech, 11368-1-AP), POT1 (1:1000, Novus Biologicals, NB500-176), HA-tag (1:1000, Cell Signalling Technology, 3724s), DYKDDDDK Tag (1:1000, Cell Signalling Technology, 14793s), H3 (1:1000, Cell Signalling Technology, D5A7), GAPDH (1:1000, Proteintech, 60004-1-Ig) followed by secondary antibody for 1 h with a 1:2000 dilution of IgG Goat Anti-Mouse HRP (Proteintech, SA00001-1) or IgG Goat Anti- Rabbit HRP (Proteintech, SA00001-2).

Techniques: Plasmid Preparation, Western Blot, Isolation, Transmission Assay

Journal: eBioMedicine

Article Title: Modified hTERT treatment ameliorates pressure overload-induced heart failure

doi: 10.1016/j.ebiom.2026.106203

Figure Lengend Snippet: JV101 blocked DDR-p53-TFAM axis and inflammation response in Ang II-induced hiPSC-CMs. (A) Expression levels of inflammatory cytokines in hiPSC-CMs were determined by RT-qPCR (n = 6). (B) Representative micrographs of hiPSC-CMs displaying Telomere (red), 53BP1 (green) and DAPI (blue). (C) Quantification of 53BP1-telomere co-localisation (n = 6). (D) Cell lysate isolated from Ang II + Vector, Ang II + JV101 and control hiPSC-CMs were immunoblotted for p-p53(Ser15), p53, PGC-1a and TFAM. (E–G) Representative transmission electron micrographs of left ventricular myocardium of the indicated study groups. Quantitative analyses of cristae number (F) and mitochondria density (G) in hiPSC-CMs. (H) Quantification of mitochondrial DNA (mtDNA) copy numbers in different groups (n = 6). (I) Intracellular ATP content of different groups (n = 6). Data are represented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. Statistical differences among groups were analysed by one-way ANOVA followed by Tukey's multiple comparisons test.

Article Snippet: The primary antibodies: histone H 2 AX (1:1000, Cell Signalling Technology, 7631S), γH 2 AX (1:1000, Cell Signalling Technology, 9718S), 53BP1(1:1000, Cell Signalling Technology, 4937S), p53 (1:1000, Cell Signalling Technology, 2524S), Phospho-p53 (Ser15) (1:1000, Cell Signalling Technology, 9284s), PGC-1α (1:1000, Santa Cruz, sc-517380), TFAM (1:1000, Proteintech, 22586-1-AP), TRF1 (1:1000, Abcam, ab192629), RAP1 (1:1000, Santa Cruz, sc-53434), TIN2 (1:1000, Proteintech, 11368-1-AP), POT1 (1:1000, Novus Biologicals, NB500-176), HA-tag (1:1000, Cell Signalling Technology, 3724s), DYKDDDDK Tag (1:1000, Cell Signalling Technology, 14793s), H3 (1:1000, Cell Signalling Technology, D5A7), GAPDH (1:1000, Proteintech, 60004-1-Ig) followed by secondary antibody for 1 h with a 1:2000 dilution of IgG Goat Anti-Mouse HRP (Proteintech, SA00001-1) or IgG Goat Anti- Rabbit HRP (Proteintech, SA00001-2).

Techniques: Expressing, Quantitative RT-PCR, Isolation, Plasmid Preparation, Control, Transmission Assay

Journal: International Journal of Oncology

Article Title: Overcoming acquired doxorubicin resistance of ovarian carcinoma cells by verapamil-mediated promotion of DNA damage-driven cytotoxicity

doi: 10.3892/ijo.2026.5861

Figure Lengend Snippet: Effect of Topo II inhibitors on DNA damage formation and activation of DDR-related mechanisms in A2780 and A2780ADR cells. (A) At 24 h after treatment of logarithmically growing cells with the indicated concentrations of Doxo or Eto, the number of nuclear γH2AX-foci, 53BP1-foci, γH2AX/53BP1 co-localized foci and γH2AX pan-stained cells was analyzed. The upper part of the figure shows representative images (total magnification, ×1,000). Quantitative data depicted in the histogram are the mean ± SD from n=3 independent experiments with each five images being analyzed per experimental condition. * P≤0.05; ** P≤0.01; *** P≤0.001 (A2780 vs. A2780ADR); # P≤0.05; ## P≤0.01; ### P≤0.001 (treated vs. untreated control). Control experiments performed by use of 1st or 2nd antibody only or no antibody at all did not interfere with the signal of main interest (that is, nuclear foci; data not shown). (B) Logarithmically growing cells were treated with the indicated concentrations of Doxo for 24 or 72 h. Afterwards, the protein expression of DDR-related factors was analyzed by western blotting using EKR2 protein expression as loading control. Doxo, doxorubicin; Eto, etoposide; p-, phosphorylated; Chk1/2, checkpoint kinase 1/2; ERK2, extracellular regulated kinase; γH2AX, Ser139 phosphorylated histone H2AX; 2; Kap1, KRAB-associated protein 1; PARP, poly (ADP-ribose) polymerase; p21, cyclin-dependent kinase inhibitor 1; p53, tumor suppressor p53.

Article Snippet: The following primary antibodies were used: ATP7A (cat. no. PA5-103110; Invitrogen; Thermo Fisher Scientific; Inc.), Caspase-7 cleaved (Asp198; cat. no. 9491S; Cell Signaling Technology, Inc.), Chk1phospho (Ser345; cat. no. 2341, Cell Signaling Technology, Inc.), Chk2phosphoT68 (Y171; cat. no. ab32148; Abcam), CTR1/SLC31A1 (EPR7936; cat. no. ab129067; Abcam), Cyclin B1 (cat. no. 4138, Cell Signaling Technology, Inc.), ERK2 [PA5-32396, Invitrogen/Thermo Fisher Scientific; Carlsbad, USA], Galactosidase beta (E2U2I) (cat. no. 27198, Cell Signaling Technology, Inc.), GAPDH (14C10; cat. no. 2118S; Cell Signaling Technology, Inc.), H2AX phospho (Ser139, cloneJBW301; cat. no. 05-636, Merck KGaA), MDR1/ABCB1 (D3H1Q; cat. no. 12683; Cell Signaling Technology, Inc.), OCT2 (cat. no. MBS9600162, Biozol Diagnostics Vertrieb GmbH), P16 (F-12; cat. no. sc-1661; Santa Cruz Biotechnology, Inc.), p21 (C-19; cat. no. sc-397; Santa Cruz Biotechnology, Inc.), P53 phospho (S15; cat. no. 9284S; Cell Signaling Technology, Inc.), PARP (cat. no. 9542S, Cell Signaling Technology, Inc.), Rad51 (cat. no. ab63801; Abcam), RPA32 phospho (S4/S8; cat. no. ICH-00422; Bethyl Laboratories Inc.), TopBP1 (D8G4L; cat. no. 14342, Cell Signaling Technology, Inc.), Topoisomerase II alpha (D10G9; cat. no. 12286, Cell Signaling Technology, Inc.).

Techniques: Activation Assay, Staining, Control, Expressing, Western Blot

Journal: International Journal of Oncology

Article Title: Overcoming acquired doxorubicin resistance of ovarian carcinoma cells by verapamil-mediated promotion of DNA damage-driven cytotoxicity

doi: 10.3892/ijo.2026.5861

Figure Lengend Snippet: Influence of combined treatment of A2780ADR cells with Doxo and selected inhibitors on mechanism of the DDR and mRNA expression of selected susceptibility-related genes. (A) Logarithmically growing A2780ADR cells were co-treated with the indicated concentrations of Doxo and selected pharmacological inhibitors (concentrations see ) for 24 or 72 h. Afterwards, the protein expression of DDR-related factors was analyzed by western blotting. For loading control, blots were reprobed with ERK2 antibody. (B) Reverse transcription-quantitative PCR of the mRNA expression of selected factors known to contribute to different mechanisms of drug sensitivity. Data shown are mean ± SD from triplicate determinations as described in methods. Relative mRNA level in untreated A2780ADR cells was set to 1.0. Doxo, doxorubicin; DDR, DNA damage response; p-, phosphorylated; nd, not detectable; Bax, Bcl-2 associated protein X; Bcl-2, B-cell lymphoma; BBC3, Bcl-2 binding component 2; BRCA1, 2, breast cancer associated gene 1,2; Cl casp-7, cleaved caspase 7; Chk, checkpoint kinase; CXCL8, chemokine ligand 8 (interleukin 8); p21, CDK inhibitor 1; p16, CDK inhibitor 2; CDKN1A/2A, cyclin dependent kinae inhibitor 1A/2A; CCNB1, Cyclin B1; b-Gal, beta-galactosidase; FASL, FAS ligand; FASR, FAS receptor; GADD, growth arrest and DNA damage inducible GPX1, glutathione peroxidase 1; GSTM1, glutathione S-transferase 1; HMOX1, heme oxygenase 1; γH2AX, Ser139 phosphorylated histone H2AX; p53, tumor suppressor p53; PARP, poly (ADP-ribose) polymerase; PCNA-proliferating cell nuclear antigen; PGC1A, PPARG coactivator 1; PPARGC1A, peroxisome proliferator-activated receptor gamma coactivator 1-alpha; RAD51, radiation damage gene 51; RPAreplication protein A; SOD1, superoxide dismutase 1; Ver, verapamil.

Article Snippet: The following primary antibodies were used: ATP7A (cat. no. PA5-103110; Invitrogen; Thermo Fisher Scientific; Inc.), Caspase-7 cleaved (Asp198; cat. no. 9491S; Cell Signaling Technology, Inc.), Chk1phospho (Ser345; cat. no. 2341, Cell Signaling Technology, Inc.), Chk2phosphoT68 (Y171; cat. no. ab32148; Abcam), CTR1/SLC31A1 (EPR7936; cat. no. ab129067; Abcam), Cyclin B1 (cat. no. 4138, Cell Signaling Technology, Inc.), ERK2 [PA5-32396, Invitrogen/Thermo Fisher Scientific; Carlsbad, USA], Galactosidase beta (E2U2I) (cat. no. 27198, Cell Signaling Technology, Inc.), GAPDH (14C10; cat. no. 2118S; Cell Signaling Technology, Inc.), H2AX phospho (Ser139, cloneJBW301; cat. no. 05-636, Merck KGaA), MDR1/ABCB1 (D3H1Q; cat. no. 12683; Cell Signaling Technology, Inc.), OCT2 (cat. no. MBS9600162, Biozol Diagnostics Vertrieb GmbH), P16 (F-12; cat. no. sc-1661; Santa Cruz Biotechnology, Inc.), p21 (C-19; cat. no. sc-397; Santa Cruz Biotechnology, Inc.), P53 phospho (S15; cat. no. 9284S; Cell Signaling Technology, Inc.), PARP (cat. no. 9542S, Cell Signaling Technology, Inc.), Rad51 (cat. no. ab63801; Abcam), RPA32 phospho (S4/S8; cat. no. ICH-00422; Bethyl Laboratories Inc.), TopBP1 (D8G4L; cat. no. 14342, Cell Signaling Technology, Inc.), Topoisomerase II alpha (D10G9; cat. no. 12286, Cell Signaling Technology, Inc.).

Techniques: Expressing, Western Blot, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Binding Assay

Journal: bioRxiv

Article Title: Single-cell full-length transcriptome of human lung reveals genetic effects on isoform regulation beyond gene-level expression

doi: 10.64898/2026.03.27.714873

Figure Lengend Snippet: ( A ) Pie chart presents the fraction of GWAS-colocalized eIsoforms with an eQTL-colocalizing isoQTL for a matched eGene in the same cell type. ( B ) Pie chart presents the fraction of TWAS significant isoforms overlapping TWAS genes from cell-barcode matched short-read eQTL data ( C ) isoQTL or eQTL colocalization with lung cancer GWAS at the PPIL6 locus in multiciliated cells. SNPs are color-coded based on the LD R 2 (1000 Genomes, EAS, phase 3) with isoQTL lead SNP, rs12528822 (purple diamond). ( D ) Association between the genotype of lead isoQTL rs12528822 and the normalized expression of PPIL6-207 (top panel) and TALONT003040002 (bottom panel). ( E ) Association between the genotype of lead isoQTL rs12528822 and the proportion of PPIL6-207 (top panel) and TALONT003040002 (bottom panel) over all PPIL6 isoforms. P -value is calculated by linear regression with the same covariates used in isoQTL mapping adjusted. Allele C of rs12528822 is the risk-associated allele for lung cancer. The violins and boxes are colored by genotype, and the grey line shows the trend of association. Center lines show the medians; the box indicates the middle of 50% of data; whiskers extend 1.5 times the interquartile range from the 25th and 75th percentiles; outliers are represented by dots; density of normalized expression is represented by the width of violin shape. ( F , H ) Representative histograms of DNA damage marker γH2AX ( F ) or p-p53 ( H ) for overproduced PPIL6 isoforms or HPS4 in GFP + transfected cells from the same experimental batch. ( G,I ) Bar plots showed the DNA damage level (i.e., γH2AX or p-p53) normalized to the median intensity of GFP + HPS4-overproducing MRC5-SV40 cells. Error bars show the mean±standard error. Black dots show the individual level of normalized value of two replicates from three experiments (n = 6). One-way ANOVA is used to test the differences across groups, * represents adjusted p- value < 0.05 by post hoc Tukey’s test. All summary statistics are in Table S11 .

Article Snippet: Antibody staining was performed using anti–phospho–histone H2A.X (Ser139) antibody (clone JBW301; 1:750; Sigma–Aldrich, 05-636-25UG) and p-p53 (Ser15) mouse monoclonal antibody (clone 16G8; 1:1000; Cell Signaling Technology, 9286), and Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa FluorTM 647 (1:1000 dilution; Invitrogen, A-21235).

Techniques: Expressing, Marker, Transfection