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: Cell Cycle

Article Title: Efficacy of poly (ADP-ribose) polymerase inhibitor olaparib against head and neck cancer cells: Predictions of drug sensitivity based on PAR–p53–NF-κB interactions

doi: 10.1080/15384101.2016.1235104

Figure Lengend Snippet: Olaparib reduced the viability of AMC-HN4 cells via the suppression of NF-κB signaling. (A) A comet assay and (B) γH2AX immunofluorescence assay were performed 72 h after olaparib treatment to identify DNA damage. A relatively higher level of DNA damage was observed in HN9-cisR cells; however, olaparib also induced slight DNA damage in olaparib-resistant HN4-cisR cells. Magnification: × 100 (comet assay); × 400 (γH2AX). (C) Western blot analysis in HN9-cisR cells according to changes in olaparib doses. Olaparib induced pATM and 53BP1 activation in a dose-dependent manner in HN9-cisR cells. (D) Western blot analyses of HN4 and HN9-cisR cells according to the indicated time points after 20-μM olaparib treatment. NF-κB was more strongly expressed in HN4 cells, compared to HN9-cisR cells, and this expression decreased significantly in a time-dependent manner after olaparib treatment. (E) ATM, 53BP1, RAD51, and γH2AX immunofluorescence assays to detect DNA damage 72 h after olaparib treatment. Magnification: × 400.

Article Snippet: The following primary antibodies were used: PAR (Enzo Life Sciences, Farmingdale, NY, USA); ATM, NF-κB p65 (NF-κB), p53, and RAD51 (Santa Cruz Biotechnology, Dallas, TX, USA); apoptosis-inducing factor (AIF), BAX, γH2AX, PARP-1 with cleaved PARP, p21WAF1/CIP1 (p21); BAY 11-7089 and betulinic acid (Tocris Bioscience, Bristol, UK), and phospho-p53-Ser15 (pp53) (Cell Signaling Technology, Danvers, MA, USA). β-actin (Sigma-Aldrich) was used as the loading control; all antibodies were used at dilutions ranging from 1:250 to 1:5,000.

Techniques: Single Cell Gel Electrophoresis, Immunofluorescence, Western Blot, Activation Assay, Expressing