Journal: Nature communications

Article Title: Cdkn1b overexpression in adult mice alters the balance between genome and tissue aging

doi: 10.1038/ncomms3626

Figure Lengend Snippet: Panel A shows western blots for pp53(ser18) (a full-length image of this blot is shown in Supplementary Figure 10, panel B) and actin in lung, intra-abdominal fat (IA Fat), and skin of R26-M2rtTA transgenic mice that either carry or do not carry the TRE-Tight-Cdkn1b transgene as indicated in the figure where, starting at two months of age, mice were continuously treated with 1 mg/ml doxycycline for two months and assessed at 4 months of age. Additionally, Cdkn1b expression in the same tissues from a control 24 month old mouse is included for comparison. Arrows indicate the positions of size markers in kDa. Panels B and C show immune-fluorescence staining for γ-H2AX in epidermal sections of control (B) and Cdkn1b over-expressing (C) mice. Panels D and E show immune-fluorescence staining for TP53BP1 in epidermal sections of control (D) and Cdkn1b over-expressing (E) mice. Panels F and G demonstrate reduced frequency of γ-H2AX (n = 9, error bars indicate s.d., ρ = 0.04, two-tailed t-test) and TP53BP1 (n = 10, error bars indicate s.d , ρ = 3.7 × 10 −5 , two-tailed t-test) foci, respectively, in epidermal sections from Cdkn1b over-expressing mice relative to controls. Panel H shows SA-β galactosidase staining of intra-abdominal fat (IAT) for R26-M2rtTA control and R26-M2rtTA;TRE-Tight-Cdkn1b transgenic mice that were treated with 1 mg/ml doxycycline either from birth through 3 months of age (left panels) or starting at 2 months of age and through 8 months of age (right panels). Bars = 5 mm. Quantitative assessment of SA-β galactosidase activity was made using ONPG as a substrate and results from this assay are shown below each panel. Additionally, the weight of total IAT recovered from each animal is given in grams (g). Qualitatively similar SA-β galactosidase staining was observed in a two additional sets of control and R26-M2rtTA;TRE-Tight-Cdkn1b mice treated with 0.25 mg/ ml doxycycline from months 2–7 although quantitative measures were not performed.

Article Snippet: Following immunoblotting (western blot) Cdkn1b, pp53(ser18), and actin were detected using anti-p27(C-19) (1/1000, Santa Cruz Biotechnology Cat No sc-528), anti-pp53(ser18) (1/1000, Cell Signaling Cat No 9284S), or anti-actin (1/1000, Sigma Cat No A2066) primary antibodies and HRP conjugated anti-rabbit secondary antibody (1/2000, Mllipore AP132P GtxRb IgG (H+L) HRP).

Techniques: Western Blot, Transgenic Assay, Expressing, Control, Comparison, Fluorescence, Staining, Two Tailed Test, Activity Assay

Journal: Radiation Research

Article Title: BMS-345541 Sensitizes MCF-7 Breast Cancer Cells to Ionizing Radiation by Selective Inhibition of Homologous Recombinational Repair of DNA Double-Strand Breaks

doi: 10.1667/rr3034.1

Figure Lengend Snippet: FIG. 4. BMS enhances DNA damage response (DDR) to IR. MCF-7 cells were incubated with vehicle or 5 lM BMS for 1 h before exposure to 2 Gy IR. IR-induced phosphorylation of ATM and Chk2 were analyzed by immunofluorescent staining, and p53 phosphorylation was analyzed by Western blot at 1 and 6 h after irradiation. The cells without exposure to IR were also included as a control (CTL). Panel A: Representative photomicrographs (1003 magnifications) of phosphorylated ATM (pATM, green) and Chk2 (pChk2, red) immunofluorescent stainings and nucleic counterstaining with Hoechst-33342 (blue) are shown. Panels B and C: Average numbers (6SE) of pATM and pChk2 foci/cell are presented. ***P , 0.001 vs. vehicle. Panel D: A representative analysis of the levels of phosphorylated p53 (pp53), total p53 and b-actin in MCF- 7 cells by Western blots is shown. Similar results were also observed in two additional analyses.

Article Snippet: Western blot analysis was carried out as previously reported (3), with the following primary antibodies: anti-Rad51 (no. ab213; Abcam), Brca1 (no. ab16780; Abcam), Oct1 (no. 8157, Cell Signaling Technology, Danvers, MA), eEF2 (no. 2332, Cell Signaling Technology), p53 (no. 9282, Cell Signaling Technology), phosphop53 (pp53 [Ser15]) (no. 9286, Cell Signaling Technology), and Actin (no. SC-1616, Santa Cruz Biotechnology, Santa Cruz, CA).

Techniques: Incubation, Phospho-proteomics, Staining, Western Blot, Irradiation, Control

Journal: Cancers

Article Title: Regulation of PD-L1 Expression by SAHA-Mediated Histone Deacetylase Inhibition in Lung Cancer Cells

doi: 10.3390/cancers17172919

Figure Lengend Snippet: Mechanisms of PD-L1 regulation in lung cancer. PD-L1 expression in tumor cells is regulated at multiple levels. The p53–p21 axis can negatively influence PD-L1 transcription, while oncogenic pathways such as PI3K/AKT and JAK/STAT promote its up-regulation. Epigenetic mechanisms play a critical role: histone acetylation and methylation dynamically regulate chromatin accessibility, and DNA methylation (e.g., via DNMT1) can repress or activate PD-L1 transcription depending on context. Collectively, these mechanisms contribute to PD-L1-mediated immune evasion and tumor survival.

Article Snippet: pp53 (Phospho-p53) , CST , 12571 , 1:1000.

Techniques: Expressing, Methylation, DNA Methylation Assay

Journal: CytoJournal

Article Title: Phosphatase and tensin homolog accelerate the inflammatory injury and glycolysis of chondrocytes through TP53 protein-mediated p38MAPK pathway activation in a high-glucose environment

doi: 10.25259/Cytojournal_199_2024

Figure Lengend Snippet: TP53 is a binding protein of PTEN. (a) Potential binding proteins of PTEN. (b) Binding relationship of PTEN and TP53 was validated through Co-IP. (c ) Primary chondrocytes were transfected with pcDNA-PTEN or small interfering -PTEN for 48 h. Then, qPCR was performed to detect TP53 mRNA expression. n = 3. ✶ P < 0.05. PTEN: Phosphatase and tensin homolog, TP53: Tumor suppressor protein p53, mRNA: Messenger RNA, Co-IP: Co-immunoprecipitation, qPCR: Quantitative real-time polymerase chain reaction, pcDNA: Plasmid DNA, IP: Immunoprecipitation, IB: Immunoblotting.

Article Snippet: [ - ] The si-PTEN, siRNA against tumor suppressor protein p53 (TP53) (si-TP53), and their negative controls (scramble) were provided by Santa Cruz Biotechnology (PTEN: sense: 5'-GCA CAA GAG GCC CUA GAU UTT-3' and antisense: 5'-AAU CUA GGG CCU CUU GUG CTT-3'; TP53, sense: 5'-CUA CUU CCU GAA AAC GTT-3').

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Transfection, Expressing, Immunoprecipitation, Real-time Polymerase Chain Reaction, Plasmid Preparation, Western Blot

Journal: CytoJournal

Article Title: Phosphatase and tensin homolog accelerate the inflammatory injury and glycolysis of chondrocytes through TP53 protein-mediated p38MAPK pathway activation in a high-glucose environment

doi: 10.25259/Cytojournal_199_2024

Figure Lengend Snippet: Effects of TP53 on the injury and glycolysis of chondrocytes in an HG environment. The HG-stimulated primary chondrocytes were transfected with scramble or small interfering-TP53 for 48 h. (a) mRNA expression of TP53. (b and c) Protein expression of TP53. (d) Cell viability. (e and f) Cell apoptosis. (g) Content of IL-6 and TNF-a. (h and i) Protein expression of MMP13 and collagen II. (j) Glucose uptake (2-NBDG fluorescent probe). (k) Lactate detection kits were used to detect lactic acid content. (l and m) ROS generation. (n and o) Protein expression of HK2 and LDHA (glycolytic molecules). n = 3. ✶ P < 0.05. HG: High glucose, TP53: Tumor suppressor protein p53, IL-6: Interleukin-6, TNF-a: Tumor necrosis factor-alpha, MMP13: Matrix metalloproteinase 13, ROS: Reactive oxygen species, HK2: Hexokinase2, LDHA: Lactate dehydrogenase A, mRNA: Messenger RNA, IL: Interleukin, DAPI: 4’,6-diamidino-2-phenylindole, FITC: Fluorescein isothiocyanate, PI: Propidium iodide. Scale bar = 25 μm (400×).

Article Snippet: [ - ] The si-PTEN, siRNA against tumor suppressor protein p53 (TP53) (si-TP53), and their negative controls (scramble) were provided by Santa Cruz Biotechnology (PTEN: sense: 5'-GCA CAA GAG GCC CUA GAU UTT-3' and antisense: 5'-AAU CUA GGG CCU CUU GUG CTT-3'; TP53, sense: 5'-CUA CUU CCU GAA AAC GTT-3').

Techniques: Transfection, Expressing

Journal: CytoJournal

Article Title: Phosphatase and tensin homolog accelerate the inflammatory injury and glycolysis of chondrocytes through TP53 protein-mediated p38MAPK pathway activation in a high-glucose environment

doi: 10.25259/Cytojournal_199_2024

Figure Lengend Snippet: PTEN affects the injury and glycolysis of chondrocytes by upregulating TP53 in an HG environment. (a and b) Protein expression of TP53. (c) Cell viability. (d and e) Cell apoptosis. (f) IL-6 secretion levels. (g and h) MMP13 protein expression. (i) Glucose uptake. (j) Lactate detection kits were used to detect lactic acid content. (k and l) ROS generation. (m and n) Glycolytic molecules HK2 protein expression. n = 3. ✶ P < 0.05. HG: High glucose, PTEN: Phosphatase and tensin homolog, TP53: Tumor suppressor protein p53, IL-6: Interleukin-6, MMP13: Matrix metalloproteinase 13, ROS: Reactive oxygen species, HK2: Hexokinase2, DAPI: 4’,6-diamidino-2-phenylindole, PI: Propidium iodide. Scale bar = 25 μm (400×).

Article Snippet: [ - ] The si-PTEN, siRNA against tumor suppressor protein p53 (TP53) (si-TP53), and their negative controls (scramble) were provided by Santa Cruz Biotechnology (PTEN: sense: 5'-GCA CAA GAG GCC CUA GAU UTT-3' and antisense: 5'-AAU CUA GGG CCU CUU GUG CTT-3'; TP53, sense: 5'-CUA CUU CCU GAA AAC GTT-3').

Techniques: Expressing

Journal: CytoJournal

Article Title: Phosphatase and tensin homolog accelerate the inflammatory injury and glycolysis of chondrocytes through TP53 protein-mediated p38MAPK pathway activation in a high-glucose environment

doi: 10.25259/Cytojournal_199_2024

Figure Lengend Snippet: PTEN affects injury and glycolysis of chondrocytes by up-regulating TP53 to activate the p38MAPK pathway in an HG environment. SB203580, a p38 pathway inhibitor. (a and b) Phosphorylation level of p38 protein. (c) TNF-a secretion level. (d and e) Cell apoptosis. (f) Glucose uptake. (g) Lactic acid content. (h and i) ROS generation. (j and k) Glycolytic molecule LDHA protein expression. (l and m) Collagen II protein expression. n = 3. ✶ P <0.05. HG: High glucose, PTEN: Phosphatase and tensin homolog, TP53: Tumor suppressor protein p53, TNF-a: Tumor necrosis factor-alpha, ROS: Reactive oxygen species, LDHA: Lactate dehydrogenase A, p38MAPK: p38 mitogen-activated protein kinase, FITC: Fluorescein isothiocyanate, PI: Propidium iodide. Scale bar = 25 μm (400×).

Article Snippet: [ - ] The si-PTEN, siRNA against tumor suppressor protein p53 (TP53) (si-TP53), and their negative controls (scramble) were provided by Santa Cruz Biotechnology (PTEN: sense: 5'-GCA CAA GAG GCC CUA GAU UTT-3' and antisense: 5'-AAU CUA GGG CCU CUU GUG CTT-3'; TP53, sense: 5'-CUA CUU CCU GAA AAC GTT-3').

Techniques: Phospho-proteomics, Expressing

Journal: CytoJournal

Article Title: Phosphatase and tensin homolog accelerate the inflammatory injury and glycolysis of chondrocytes through TP53 protein-mediated p38MAPK pathway activation in a high-glucose environment

doi: 10.25259/Cytojournal_199_2024

Figure Lengend Snippet: Effects of PTEN on the inflammatory injury and glycolysis of cartilage tissue in rats with DOA. (a and b) Representative images of immunohistochemistry for PTEN and TP53. Scale bar = 50 μm (×200) and 25 μm (×400). (c and d) Phosphorylation level of p38MAPK protein. (e and f) Protein expression of collagen II. (e and g) Glycolytic molecule HK-2 protein expression. (h) IL-6 secretion level. (i) Glucose uptake. (j) Lactic acid content. (k) Representative images of the hematoxylin and eosin staining of the knee cartilage tissue. ✶ P < 0.05. DOA: Diabetic osteoarthritis, PTEN: Phosphatase and tensin homolog, TP53: Tumor suppressor protein p53, IL-6: Interleukin-6, HK2: Hexokinase2, p38MAPK: p38 mitogen-activated protein kinase. Scale bar = 50 μm and 25 μm.

Article Snippet: [ - ] The si-PTEN, siRNA against tumor suppressor protein p53 (TP53) (si-TP53), and their negative controls (scramble) were provided by Santa Cruz Biotechnology (PTEN: sense: 5'-GCA CAA GAG GCC CUA GAU UTT-3' and antisense: 5'-AAU CUA GGG CCU CUU GUG CTT-3'; TP53, sense: 5'-CUA CUU CCU GAA AAC GTT-3').

Techniques: Immunohistochemistry, Phospho-proteomics, Expressing, Staining

Journal: Cancers

Article Title: Tipping Growth Inhibition into Apoptosis by Combining Treatment with MDM2 and WIP1 Inhibitors in p53 WT Uterine Leiomyosarcoma

doi: 10.3390/cancers14010014

Figure Lengend Snippet: Details of antibodies used for Western immunoblotting. BSA, Bovine serum albumin.

Article Snippet: pp53 , (E9Y4U) #82530 , Rabbit , Cell Signalling Technologies , 1:1000 , BSA.

Techniques: Western Blot