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monoclonal mouse anti p53  (Santa Cruz Biotechnology)


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    Structured Review

    Santa Cruz Biotechnology monoclonal mouse anti p53
    A mutualistic association of CDK5 and 18E6 proteins. (A) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, and 4 μg) and a fixed amount of pcDNA3.1: HA-18E6 (2 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, and 4 μg) and pcDNA3.1: His-CDK5 (2 μg). (B) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein in the HeLa cell lines. HeLa cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, 3, and 4 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein in the SAS cell lines. SAS cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, 3, and 4 μg). (C) (i). The represented immunoblot image of CDK5, E6AP, and <t>p53</t> after silencing 18E6/E7 expression. Total protein lysates were extracted from HeLa cell lines transfected with siRNA against control (siCtrl) or 18 E6 and E7 (si18 E6/E7) for 72 h. (ii-v) The relative expression levels of 18E6, CDK5, p53, and E6AP relative to β-actin and analyzed using ImageJ and GraphPad Prism (n = 3). All data were presented as means ± standard error of the mean (SEM). (∗, P < 0.05; ∗∗, P < 0.01). (D) The represented immunoblot of CDK5, pCDK5 and HPV 18E6 proteins in the stable expression of HPV 18 in the HGK12 cell line (HPV-null primary keratinocytes cell line), HC: Treatment with 0.2 μM CP681301; LC: Treatment with 0.1 μM CP681301.
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    Images

    1) Product Images from "HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer"

    Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

    Journal: Tumour Virus Research

    doi: 10.1016/j.tvr.2026.200339

    A mutualistic association of CDK5 and 18E6 proteins. (A) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, and 4 μg) and a fixed amount of pcDNA3.1: HA-18E6 (2 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, and 4 μg) and pcDNA3.1: His-CDK5 (2 μg). (B) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein in the HeLa cell lines. HeLa cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, 3, and 4 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein in the SAS cell lines. SAS cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, 3, and 4 μg). (C) (i). The represented immunoblot image of CDK5, E6AP, and p53 after silencing 18E6/E7 expression. Total protein lysates were extracted from HeLa cell lines transfected with siRNA against control (siCtrl) or 18 E6 and E7 (si18 E6/E7) for 72 h. (ii-v) The relative expression levels of 18E6, CDK5, p53, and E6AP relative to β-actin and analyzed using ImageJ and GraphPad Prism (n = 3). All data were presented as means ± standard error of the mean (SEM). (∗, P < 0.05; ∗∗, P < 0.01). (D) The represented immunoblot of CDK5, pCDK5 and HPV 18E6 proteins in the stable expression of HPV 18 in the HGK12 cell line (HPV-null primary keratinocytes cell line), HC: Treatment with 0.2 μM CP681301; LC: Treatment with 0.1 μM CP681301.
    Figure Legend Snippet: A mutualistic association of CDK5 and 18E6 proteins. (A) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, and 4 μg) and a fixed amount of pcDNA3.1: HA-18E6 (2 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, and 4 μg) and pcDNA3.1: His-CDK5 (2 μg). (B) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein in the HeLa cell lines. HeLa cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, 3, and 4 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein in the SAS cell lines. SAS cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, 3, and 4 μg). (C) (i). The represented immunoblot image of CDK5, E6AP, and p53 after silencing 18E6/E7 expression. Total protein lysates were extracted from HeLa cell lines transfected with siRNA against control (siCtrl) or 18 E6 and E7 (si18 E6/E7) for 72 h. (ii-v) The relative expression levels of 18E6, CDK5, p53, and E6AP relative to β-actin and analyzed using ImageJ and GraphPad Prism (n = 3). All data were presented as means ± standard error of the mean (SEM). (∗, P < 0.05; ∗∗, P < 0.01). (D) The represented immunoblot of CDK5, pCDK5 and HPV 18E6 proteins in the stable expression of HPV 18 in the HGK12 cell line (HPV-null primary keratinocytes cell line), HC: Treatment with 0.2 μM CP681301; LC: Treatment with 0.1 μM CP681301.

    Techniques Used: Western Blot, Transfection, Expressing, Control

    CP681301 treatment destabilized E6, reduced E6AP and rescued p53. (A). HeLa and C33A cell lines were treated with CP681301 at the concentrations of 0.6 μM and 1.5 μM for 24 h. The protein extracts were subjected to Western blotting for the detection of CDK5, 18E6, E6AP, p53, and β-actin proteins. (B). CP681301 disrupted the 18E6-E6AP protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and Flag-E6AP protein for 2h. After extensive washing, the bound E6AP protein was detected via Western blotting using an anti-Flag antibody. The immunoblot (IB) on the upper panel shows the interaction of E6AP with GST-18E6, while the lower panel shows the Ponceau S stained of the blot.
    Figure Legend Snippet: CP681301 treatment destabilized E6, reduced E6AP and rescued p53. (A). HeLa and C33A cell lines were treated with CP681301 at the concentrations of 0.6 μM and 1.5 μM for 24 h. The protein extracts were subjected to Western blotting for the detection of CDK5, 18E6, E6AP, p53, and β-actin proteins. (B). CP681301 disrupted the 18E6-E6AP protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and Flag-E6AP protein for 2h. After extensive washing, the bound E6AP protein was detected via Western blotting using an anti-Flag antibody. The immunoblot (IB) on the upper panel shows the interaction of E6AP with GST-18E6, while the lower panel shows the Ponceau S stained of the blot.

    Techniques Used: Western Blot, Inhibition, Pull Down Assay, Incubation, Purification, Staining



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    A mutualistic association of CDK5 and 18E6 proteins. (A) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, and 4 μg) and a fixed amount of pcDNA3.1: HA-18E6 (2 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, and 4 μg) and pcDNA3.1: His-CDK5 (2 μg). (B) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein in the HeLa cell lines. HeLa cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, 3, and 4 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein in the SAS cell lines. SAS cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, 3, and 4 μg). (C) (i). The represented immunoblot image of CDK5, E6AP, and p53 after silencing 18E6/E7 expression. Total protein lysates were extracted from HeLa cell lines transfected with siRNA against control (siCtrl) or 18 E6 and E7 (si18 E6/E7) for 72 h. (ii-v) The relative expression levels of 18E6, CDK5, p53, and E6AP relative to β-actin and analyzed using ImageJ and GraphPad Prism (n = 3). All data were presented as means ± standard error of the mean (SEM). (∗, P < 0.05; ∗∗, P < 0.01). (D) The represented immunoblot of CDK5, pCDK5 and HPV 18E6 proteins in the stable expression of HPV 18 in the HGK12 cell line (HPV-null primary keratinocytes cell line), HC: Treatment with 0.2 μM CP681301; LC: Treatment with 0.1 μM CP681301.

    Journal: Tumour Virus Research

    Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

    doi: 10.1016/j.tvr.2026.200339

    Figure Lengend Snippet: A mutualistic association of CDK5 and 18E6 proteins. (A) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, and 4 μg) and a fixed amount of pcDNA3.1: HA-18E6 (2 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein. HEK 293 cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, and 4 μg) and pcDNA3.1: His-CDK5 (2 μg). (B) (i). The represented immunoblot image of CDK5 upregulates the level of 18E6 protein in the HeLa cell lines. HeLa cells were transfected with an increasing amount of pcDNA3.1: His-CDK5 (1, 2, 3, and 4 μg). (ii). The represented immunoblot image of 18E6 upregulates the level of CDK5 protein in the SAS cell lines. SAS cells were transfected with an increasing amount of pcDNA3.1: HA-18E6 (1, 2, 3, and 4 μg). (C) (i). The represented immunoblot image of CDK5, E6AP, and p53 after silencing 18E6/E7 expression. Total protein lysates were extracted from HeLa cell lines transfected with siRNA against control (siCtrl) or 18 E6 and E7 (si18 E6/E7) for 72 h. (ii-v) The relative expression levels of 18E6, CDK5, p53, and E6AP relative to β-actin and analyzed using ImageJ and GraphPad Prism (n = 3). All data were presented as means ± standard error of the mean (SEM). (∗, P < 0.05; ∗∗, P < 0.01). (D) The represented immunoblot of CDK5, pCDK5 and HPV 18E6 proteins in the stable expression of HPV 18 in the HGK12 cell line (HPV-null primary keratinocytes cell line), HC: Treatment with 0.2 μM CP681301; LC: Treatment with 0.1 μM CP681301.

    Article Snippet: The transferred membranes were incubated overnight at 4 °C with the following primary antibodies: monoclonal rabbit anti-HA (Cell Signaling Technology), monoclonal mouse anti-p53 (Santa Cruz), monoclonal mouse anti-HPV18E6 (Santa Cruz), monoclonal mouse anti-CDK5 (Santa Cruz), and monoclonal mouse anti-β-actin (Santa Cruz).

    Techniques: Western Blot, Transfection, Expressing, Control

    CP681301 treatment destabilized E6, reduced E6AP and rescued p53. (A). HeLa and C33A cell lines were treated with CP681301 at the concentrations of 0.6 μM and 1.5 μM for 24 h. The protein extracts were subjected to Western blotting for the detection of CDK5, 18E6, E6AP, p53, and β-actin proteins. (B). CP681301 disrupted the 18E6-E6AP protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and Flag-E6AP protein for 2h. After extensive washing, the bound E6AP protein was detected via Western blotting using an anti-Flag antibody. The immunoblot (IB) on the upper panel shows the interaction of E6AP with GST-18E6, while the lower panel shows the Ponceau S stained of the blot.

    Journal: Tumour Virus Research

    Article Title: HPV18E6 and CDK5 virus-host interaction is a prospective therapeutic target for HPV-positive cervical cancer

    doi: 10.1016/j.tvr.2026.200339

    Figure Lengend Snippet: CP681301 treatment destabilized E6, reduced E6AP and rescued p53. (A). HeLa and C33A cell lines were treated with CP681301 at the concentrations of 0.6 μM and 1.5 μM for 24 h. The protein extracts were subjected to Western blotting for the detection of CDK5, 18E6, E6AP, p53, and β-actin proteins. (B). CP681301 disrupted the 18E6-E6AP protein complex. The inhibition of CP681301 was detected using the GST-pull down assay, where CP681301 was incubated with the purified GST-18E6 protein and Flag-E6AP protein for 2h. After extensive washing, the bound E6AP protein was detected via Western blotting using an anti-Flag antibody. The immunoblot (IB) on the upper panel shows the interaction of E6AP with GST-18E6, while the lower panel shows the Ponceau S stained of the blot.

    Article Snippet: The transferred membranes were incubated overnight at 4 °C with the following primary antibodies: monoclonal rabbit anti-HA (Cell Signaling Technology), monoclonal mouse anti-p53 (Santa Cruz), monoclonal mouse anti-HPV18E6 (Santa Cruz), monoclonal mouse anti-CDK5 (Santa Cruz), and monoclonal mouse anti-β-actin (Santa Cruz).

    Techniques: Western Blot, Inhibition, Pull Down Assay, Incubation, Purification, Staining

    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.

    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

    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.

    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

    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.

    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

    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.

    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