Journal: International Journal of Nanomedicine

Article Title: Activation of Erk and p53 regulates copper oxide nanoparticle-induced cytotoxicity in keratinocytes and fibroblasts

doi: 10.2147/IJN.S67688

Figure Lengend Snippet: p53 was involved in CuONP-induced cytotoxicity in HaCaT and MEF cells. Notes: Western blot analysis of p53 and p-p53 in CuONP-treated HaCaT ( A ) and MEF ( B ) cells. The effects of 1 μmol/L p53 transcriptional inhibitor (cyclic PFT-α) on CuONP-induced viability loss in HaCaT ( C ) and MEF ( E ) cells with CuONP treatment for 24 hours. The effects of 10 μmol/L cyclic PFT-α on CuONP-induced viability loss in HaCaT cells with CuONP treatment for 6 hours ( D ) and in MEF cells with CuONP treatment for 3 hours ( F ). The effects of 30 μmol/L p53 stabilizer (nutlin-3α) on CuONP-induced viability loss in HaCaT ( G ) and MEF ( H ) cells with CuONP treatment for 24 hours. The data are presented as the mean ± standard deviation (n>3). ** P <0.01 versus vehicle control. ## P <0.01 versus CuONP group. !! P <0.01 versus 1 μmol/L cyclic PFT-α group. && P <0.01 versus 10 μmol/L cyclic PFT-α group. ^^ P <0.01 versus nutlin-3α group. Abbreviations: CuONP, copper oxide nanoparticles; MEF, mouse embryonic fibroblasts; PFT-α, pifithrin-α.

Article Snippet: Primary antibodies against cyclin A, cyclin B1, p53, phosphorylated c-Jun N-terminal kinase (JNK), JNK, p-p38, p38, Erk, and cyclic pifithrin-α (PFT-α) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).

Techniques: Western Blot, Standard Deviation, Control

Journal: International Journal of Nanomedicine

Article Title: Activation of Erk and p53 regulates copper oxide nanoparticle-induced cytotoxicity in keratinocytes and fibroblasts

doi: 10.2147/IJN.S67688

Figure Lengend Snippet: Cytotoxicity of CuONP in primary keratinocytes. Notes: Viability loss in primary keratinocytes with CuONP treatment for 24 hours ( A ). The effects of Erk inhibitor (U0126, 5 μmol/L), p53 transcriptional inhibitor (cyclic PFT-α, 1 μmol/L), and p53 stabilizer (nutlin-3α, 30 μmol/L) on viability loss in primary keratinocytes with 60 μg/mL CuONP treatment for 24 hours ( B ). The data are presented as the mean ± standard deviation (n>3). ** P <0.01 versus vehicle control. ## P <0.01 versus CuONP group. !! P <0.01 versus U0126 group. ^^ P <0.01 versus cyclic PFT-α group. Abbreviations: CuONP, copper oxide nanoparticles; PFT-α, pifithrin-α.

Article Snippet: Primary antibodies against cyclin A, cyclin B1, p53, phosphorylated c-Jun N-terminal kinase (JNK), JNK, p-p38, p38, Erk, and cyclic pifithrin-α (PFT-α) were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA).

Techniques: Standard Deviation, Control

Journal: Clinical and Translational Medicine

Article Title: Oxidative stress induces monocyte‐to‐myofibroblast transdifferentiation through p38 in pancreatic ductal adenocarcinoma

doi: 10.1002/ctm2.41

Figure Lengend Snippet: The p53 stabilizer nutlin‐3 induced MMT through ROS generation but not through the p53 transcription/mitochondria‐dependent signaling pathway. A, RNA‐seq analysis of monocytes obtained from two healthy donors (D72 and D73), and treated or not treated with 100 µM H 2 O 2 for 7 days. The heatmap shows hierarchical clustering of mRNA levels of genes in monocytes treated as indicated; the volcano plot shows differentially expressed genes plotted as the log2(fold change) versus the –log10( P ‐value). A total of 94 differentially expressed genes between the H 2 O 2 ‐treated cell group and the control cell group exceeded the established thresholds (–log10( P ‐value) > 1.30 and |log2‐fold change| > .585; upregulated genes are shown in red, and downregulated genes are shown in green). KEGG pathway analysis was performed on the upregulated genes identified by RNA‐seq. B and C, Western blot analysis of the expression of p53 and the target genes p21 and MDM2 in freshly isolated monocytes (0 day), monocytes treated with 0/50/100 µM H 2 O 2 for 3/7 days, and monocytes treated with radiation (0/4/8 Gy) for 3 days. GAPDH was used as the loading control. D and E, Western blot analysis of the expression of αSMA, p53, p21, and MDM2 in monocytes treated or not treated with nutlin‐3, H 2 O 2 , PFT‐α, or PFT‐μ. GAPDH was used as the loading control. F, Representative flow cytometric analysis of intracellular ROS levels in monocytes treated or not treated with nutin‐3 (10 µM) or the same volume of DMSO (blue, ctrl; green, DMSO; red, nutlin‐3). G, Western blot analysis of monocytes treated or not treated with 10 µM nutlin‐3 or 100U/mL catalase. GAPDH was used as the loading control. Data are representative of three experiments Abbreviations: MMT, monocyte‐to‐myofibroblast transdifferentiation; RNA‐seq, RNA sequencing; DEGs, differentially expressed genes; KEGG, Kyoto Encyclopedia of Genes and Genomes; DMSO, dimethyl sulfoxide.

Article Snippet: To inhibit p53 transcription or translocation activity, monocytes were pretreated with PFT‐α (S5791; Selleck) or PFT‐μ (S2930; Selleck), respectively.

Techniques: RNA Sequencing, Control, Western Blot, Expressing, Isolation