phosphorylated p38 mitogen activated protein kinase p p38  (Cell Signaling Technology Inc)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Hsa‐miR ‐409‐3p regulates endothelial progenitor senescence via PP2A‐P38 and is a potential ageing marker in humans

doi: 10.1111/jcmm.17691

Figure Lengend Snippet: Angiogenic activities of young human EPCs after treatment with siRNA specific to PP2A (siPP2A) and western blot analysis of young human EPCs treated with PP2A siRNA. Note that (A) transcript and (B and C) protein expression levels of PP2A were suppressed in siPP2A‐treated EPCs, accompanied by attenuated cellular activities of (D) proliferation, (E) migration and (F) tube formation. n = 3 for each bar. (G) In three different clones of EPCs, phosphorylated p38 (p‐p38) and phosphorylated JNK (p‐JNK) were upregulated after siPP2A treatment. Quantitative Analysis confirmed the activation of (H) p38 and (I) JNK signalling pathways. n = 3 for each bar. Data are mean ± SEM and analysed using unpaired Student's t test for comparison between groups. * p < 0.05, *** p < 0.001 vs. corresponding negative control (NC). Abbreviations are as in Figure

Article Snippet: The antibodies included phosphorylated p38 mitogen‐activated protein kinase (p‐p38), total p38, phosphorylated c‐Jun N‐terminal kinase (p‐JNK), total JNK, phosphorylated extracellular signal‐regulated kinase (p‐ERK) and total ERK from Cell Signalling Technology.

Techniques: Western Blot, Expressing, Migration, Clone Assay, Activation Assay, Negative Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Hsa‐miR ‐409‐3p regulates endothelial progenitor senescence via PP2A‐P38 and is a potential ageing marker in humans

doi: 10.1111/jcmm.17691

Figure Lengend Snippet: Regulation of PP2A by miR‐409‐3p and the signalling proteins in young human EPCs. PP2A was downregulated by miR‐409‐3p mimic at (A) transcript and (B and C) protein levels, compared to corresponding NC. (D) Time series analysis showed, in cells treated with the mimic, (E) PP2A was downregulated for at least 72 h, while (F) p‐p38 and (G) p‐JNK were activated with peaks at 48 h. n = 5 for each bar. Data are mean ± SEM and analysed using one‐way anova followed by post hoc Fisher's least significant difference test for multiple group comparisons. * p < 0.05 vs. NC. Abbreviations are as in Figure

Article Snippet: The antibodies included phosphorylated p38 mitogen‐activated protein kinase (p‐p38), total p38, phosphorylated c‐Jun N‐terminal kinase (p‐JNK), total JNK, phosphorylated extracellular signal‐regulated kinase (p‐ERK) and total ERK from Cell Signalling Technology.

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Hsa‐miR ‐409‐3p regulates endothelial progenitor senescence via PP2A‐P38 and is a potential ageing marker in humans

doi: 10.1111/jcmm.17691

Figure Lengend Snippet: Prediction of binding site of hsa‐miR‐409‐3p for protein phosphatase 2 catalytic subunit alpha (PPP2CA) gene and examination of its product protein phosphatase 2A (PP2A). (A) Diagram showing the seed sequences of hsa‐miR‐409‐3p and its predicted binding site in the 3’‐UTR of PPP2CA gene by prediction methods including TargetScan and miRanda. (B) Luciferase reporter assays of human embryonic kidney cells 293 co‐transfected with vector, wild‐type 3’‐UTR of PPP2CA (PPP2CA‐3’UTR‐WT) or mutant 3’‐UTR of PPP2CA (PPP2CA‐3’UTR‐MUT), along with mimic negative control (NC) or miR‐409‐3p mimics showed reduced activity only in the group co‐transfected with PPP2CA‐3’UTR‐WT and miR‐409‐3p mimics. Concordantly, in senescent cells, PP2A expression was downregulated at both (C) transcript and (D and E) protein levels, with activation of (F) phosphorylated p38 (p‐p38) and (G) phosphorylated JNK (p‐JNK). n = 4 for each bar. Data are mean ± SEM and analysed using unpaired Student's t test for comparison between groups. *, p < 0.05, ***, p < 0.001 vs. corresponding control. p38, p38 mitogen‐activated protein kinase. JNK, c‐Jun N‐terminal kinase

Article Snippet: The antibodies included phosphorylated p38 mitogen‐activated protein kinase (p‐p38), total p38, phosphorylated c‐Jun N‐terminal kinase (p‐JNK), total JNK, phosphorylated extracellular signal‐regulated kinase (p‐ERK) and total ERK from Cell Signalling Technology.

Techniques: Binding Assay, Luciferase, Transfection, Plasmid Preparation, Mutagenesis, Negative Control, Activity Assay, Expressing, Activation Assay