mouse anti-bag3  (Millipore)

Journal: Journal of Cell Communication and Signaling

Article Title: miR-217-5p induces apoptosis by directly targeting PRKCI, BAG3, ITGAV and MAPK1 in colorectal cancer cells

doi: 10.1007/s12079-017-0410-x

Figure Lengend Snippet: In silico target prediction of putative target genes of miR-217

Article Snippet: These primary antibodies comprised rabbit monoclonal anti-integrin alpha V (#60896), anti-protein kinase C iota (PKCι/λ) (#2998), rabbit polyclonal anti-p44/42 MAPK (Erk1/2) (#9102) antibodies from Cell Signaling Technology (Cambridge, United Kingdom) and mouse anti-BAG3 (SAB1404732 from Sigma Aldrich).

Techniques: In Silico, Derivative Assay

Journal: Journal of Cell Communication and Signaling

Article Title: miR-217-5p induces apoptosis by directly targeting PRKCI, BAG3, ITGAV and MAPK1 in colorectal cancer cells

doi: 10.1007/s12079-017-0410-x

Figure Lengend Snippet: Localization of putative miR-217-5p binding sites in selected predicted target genes. Schematic overview of the mRNA composition of PRKCI, BAG3, ITGAV and MAPK1 and with putative miR-217-5p binding sites (blue boxes) in the 3′ untranslated region (3′ UTR) of the respective target gene mRNA (a) and the miRNA:mRNA alignment based on the data in microRNA.org (b)

Article Snippet: These primary antibodies comprised rabbit monoclonal anti-integrin alpha V (#60896), anti-protein kinase C iota (PKCι/λ) (#2998), rabbit polyclonal anti-p44/42 MAPK (Erk1/2) (#9102) antibodies from Cell Signaling Technology (Cambridge, United Kingdom) and mouse anti-BAG3 (SAB1404732 from Sigma Aldrich).

Techniques: Binding Assay

Journal: Journal of Cell Communication and Signaling

Article Title: miR-217-5p induces apoptosis by directly targeting PRKCI, BAG3, ITGAV and MAPK1 in colorectal cancer cells

doi: 10.1007/s12079-017-0410-x

Figure Lengend Snippet: PRKC1, ITGAV, BAG3 and MAPK1 as direct targets for miR-217-5p in HEK293T cells. Relative luciferase activity 3 days after co-transfection of the pMirGLO vector with the binding sites 1 to 4 and the fusion of binding sites 2 and 3 of PRKC1 (a), the binding sites of ITGAV (b) and BAG3 (c) as well with the binding sites MAPK1_1 to 6 and the fusion of binding sites 1–4 (d) with miR-217-5p mimic, miRNA inhibitor anti-miR-217-5p or non-targeting siRNA (NT). Statistical differences between means were tested using unpaired t-test [n = 3 biological and technical replicates; mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001]

Article Snippet: These primary antibodies comprised rabbit monoclonal anti-integrin alpha V (#60896), anti-protein kinase C iota (PKCι/λ) (#2998), rabbit polyclonal anti-p44/42 MAPK (Erk1/2) (#9102) antibodies from Cell Signaling Technology (Cambridge, United Kingdom) and mouse anti-BAG3 (SAB1404732 from Sigma Aldrich).

Techniques: Luciferase, Activity Assay, Cotransfection, Plasmid Preparation, Binding Assay

Journal: Journal of Cell Communication and Signaling

Article Title: miR-217-5p induces apoptosis by directly targeting PRKCI, BAG3, ITGAV and MAPK1 in colorectal cancer cells

doi: 10.1007/s12079-017-0410-x

Figure Lengend Snippet: Potential pro-apoptotic mechanisms of miR-217-5p regulating the ERK-MAPK pathway at different sites. MiR-217-5p was shown to directly downregulate PRKCI, ITGAV, BAG3 and MAPK1, connected in a signaling network modulating the ERK-MAPK pathway. Binding of extracellular matrix components to integrins comprising a β (ITGB) and α subunit such as αv (ITGAV) may activate the ERK- MAPK signaling pathway via focal adhesion kinase (FAK) activation, growth factor receptor-bound protein 2 (GRB2), and guanine nucleotide exchange factor son of sevenless (SOS), leading to the activation of the kinase cascade including KRAS, BRAF, MEK, and MAPK1 and the activation of, Phosphoinositide 3-kinase (PI3K)/Akt. These pathways culminate in the activation of survival and proliferation promoting transcription factors including c- myc, c-fos or Ets like protein 1 (Elk1), in the induction and stabilization of anti-apoptotic members (Bcl-2, Bcl-xL, Mcl-1) of the Bcl-2 protein family and the inhibition of pro-apoptotic members including the BH3-only members Bad and Bim promoting the initiation of intrinsic apoptosis via Bax and Bak. PRKCI, ITGAV, BAG3 and MAPK1 promote cell survival and proliferation by induction of transcription factors including NK-κB, AP1 or SOX2, or by induction of the ERK-MAPK pathway via Rac1

Article Snippet: These primary antibodies comprised rabbit monoclonal anti-integrin alpha V (#60896), anti-protein kinase C iota (PKCι/λ) (#2998), rabbit polyclonal anti-p44/42 MAPK (Erk1/2) (#9102) antibodies from Cell Signaling Technology (Cambridge, United Kingdom) and mouse anti-BAG3 (SAB1404732 from Sigma Aldrich).

Techniques: Binding Assay, Activation Assay, Inhibition