vegf  (Abcam)

Journal: Nucleic Acids Research

Article Title: Site-specific amino acid substitution in dodecameric peptides determines the stability and unfolding of c-MYC quadruplex promoting apoptosis in cancer cells

doi: 10.1093/nar/gky824

Figure Lengend Snippet: Conformer selectivity of the peptides for different oncogenic quadruplexes in MCF-7 cells. ( A ) pGL4.72[ hRlucCP ] vector having the inserts containing oncogenic promoter sequences ( c-MYC, BCL-2, KRAS and VEGF-A ) and upstream G-quadruplex forming elements ahead of the hRluc coding region. The promoter sequences are cloned into KpnI and HindIII restriction sites with or without the wild-type quadruplex scaffolds. hRluc, Renilla luciferase gene; hCL1 and hPEST , protein destabilizing sequences; oriC, origin of replication; AmpR, ampicillin resistance gene; SV40 (Simian virus 40 polyadenylation signal cassette), P1 and P2, promoter sequences; null, constructs having no quadruplex motif upstream the oncogene promoters; GQ, G-quadruplex forming motif. ( B ) Dual-luciferase assays. Evaluation of the promoter activity of different oncogenes (( B ) c-MYC , ( C ) BCL-2 , ( D ) KRAS and ( E ) VEGF-A ) of using the reporter plasmids with or without the wild-type quadruplex-forming sequences (Pu27-GQ, BCL-2-GQ, KRAS-GQ and VEGF-A-GQ) in MCF-7 cells. Relative promoter activity is determined by normalizing the Rluc/Fluc values to that of the cells transfected with P1-P2 promoter construct (GQ-null), having no quadruplex-forming motif. Error bars represent mean ± SE ( N = 5). Statistical differences in the luciferase activities compared to the respective GQ-null constructs of the target oncogenes used one-way ANOVA followed by Tukey–Kramer Test ( # P

Article Snippet: Santacruz Biotechnology Inc.), anti E2F1, anti VEGF-A, anti-P53, anti-APAF1, anti-Caspase 8, anti-PARP (Abcam), anti-β-actin (Mouse monoclonal, 1:10 000 dil.

Techniques: Plasmid Preparation, Clone Assay, Luciferase, Construct, Activity Assay, Transfection

Journal: Nucleic Acids Research

Article Title: Site-specific amino acid substitution in dodecameric peptides determines the stability and unfolding of c-MYC quadruplex promoting apoptosis in cancer cells

doi: 10.1093/nar/gky824

Figure Lengend Snippet: Selective arresting of c-MYC quadruplex by KR12C induce apoptotic signalling in cancer cells. ( A ) Western blot analyses of the major apoptotic markers (BCL-2, APAF-1, caspase 8 and PARP). ( B ) Estimation of protein expression level of the apoptotic markers by semi-densitometric analyses. Estimation of BCL-2 transcripts by real time PCR analyses at 5 and 10 μM KR12C concentration. ( C ) Western blot analyses of other proteins associated with the signalling cascade (c-MYC, E2F-1, VEGF-A and P53). ( D ) Quatification of protein and mRNA expression level of c-MYC, E2F-1, VEGF-A and P53 by semi-densitometric analyses and real time PCR respectively. Arrow pointing upwards and downwards denote the upregulation of the respective protein and mRNA expression level. Error bars represent mean ± SE ( N = 3). Statistical differences are determined compared to the control by two-tailed Student's t test (* P

Article Snippet: Santacruz Biotechnology Inc.), anti E2F1, anti VEGF-A, anti-P53, anti-APAF1, anti-Caspase 8, anti-PARP (Abcam), anti-β-actin (Mouse monoclonal, 1:10 000 dil.

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Concentration Assay, Two Tailed Test

Journal: Nucleic Acids Research

Article Title: Site-specific amino acid substitution in dodecameric peptides determines the stability and unfolding of c-MYC quadruplex promoting apoptosis in cancer cells

doi: 10.1093/nar/gky824

Figure Lengend Snippet: Schematic representation showing selective quadruplex interaction at c-MYC promoter by KR12C promotes apoptotic signalling in VEGF-A-BCL-2 axis in MCF-7 cells.

Article Snippet: Santacruz Biotechnology Inc.), anti E2F1, anti VEGF-A, anti-P53, anti-APAF1, anti-Caspase 8, anti-PARP (Abcam), anti-β-actin (Mouse monoclonal, 1:10 000 dil.

Techniques:

Journal: Experimental & Molecular Medicine

Article Title: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: RT-qPCR and Western blot analysis demonstrated that MBNL1-AS1 reduced the expression of KCNMA1 and cGMP-PKG signaling pathway-related genes. a RT-qPCR measured the relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. b Protein bands measured by Western blot analysis. c Western blot analysis measured the relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle cells after transfection. MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily M alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, p-PKGII phosphorylated-PKGII, VASP vasodilator-stimulated phosphoprotein, p-VASP phosphorylated-VASP, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, si small interfering, GAPDH glyceraldehyde-3-phosphate dehydrogenase, NC negative control; * p

Article Snippet: The membrane was then blocked in 5% skimmed milk at room temperature for 1 h, washed twice with PBS, and then incubated overnight at 4 °C with diluted rabbit anti-mouse primary antibodies against KCNMA1 (1:1000, MAB8589; AmyJet Scientific Inc., Wuhan, China), PKGII (1:1000, ab145063; Abcam, Inc., Cambridge, MA, USA), VASP (1:1000, ab58555; Abcam, USA), VEGF (1:1000, ab32152; Abcam, USA), Bax (1:1000, ab32503; Abcam, USA), Bcl-2 (1:1000, ab59348; Abcam, USA), Cyclin D1 (1:1000, ab134175, Abcam, USA), Cyclin D3 (1:1000, ab28283, Abcam, USA), Cdc 42 (1:1000, ab187643, Abcam, USA), caspase-3 (1:1000, ab208161, Abcam, USA), and PARP (1:1000, ab32064, Abcam, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Transfection, Negative Control

Journal: Experimental & Molecular Medicine

Article Title: Downregulation of the long noncoding RNA MBNL1-AS1 protects sevoflurane-pretreated mice against ischemia-reperfusion injury by targeting KCNMA1

doi: 10.1038/s12276-018-0133-y

Figure Lengend Snippet: MBNL1-AS1 was highly expressed, but KCNMA1 and cGMP-PKG pathway-related factors were poorly expressed in mice with I/R injury after TKA. a Relative mRNA expression of MBNL1-AS1, KCNMA1, PKGII, VASP, VEGF, Bax, and Bcl-2 in the skeletal muscle tissues of mice detected by RT-qPCR. b Protein bands observed after Western blot analysis. c Relative protein levels of KCNMA1, PKGII, VASP, p-PKGII, p-VASP, VEGF, Bax, and Bcl-2 in skeletal muscle tissues of mice evaluated by Western blot analysis. I/R ischemia-reperfusion, Sevo sevoflurane, MBNL1-AS1 muscleblind-like 1 antisense RNA 1, KCNMA1 potassium calcium-activated channel subfamily alpha 1, PKGII type II cyclic guanosine monophosphate-dependent protein kinase G, VASP vasodilator-stimulated phosphoprotein, VEGF vascular endothelial growth factor, Bcl-2 B-cell lymphoma/leukemia-2, Bax Bcl-2 associated X protein, GAPDH glyceraldehyde-3-phosphate dehydrogenase; * p

Article Snippet: The membrane was then blocked in 5% skimmed milk at room temperature for 1 h, washed twice with PBS, and then incubated overnight at 4 °C with diluted rabbit anti-mouse primary antibodies against KCNMA1 (1:1000, MAB8589; AmyJet Scientific Inc., Wuhan, China), PKGII (1:1000, ab145063; Abcam, Inc., Cambridge, MA, USA), VASP (1:1000, ab58555; Abcam, USA), VEGF (1:1000, ab32152; Abcam, USA), Bax (1:1000, ab32503; Abcam, USA), Bcl-2 (1:1000, ab59348; Abcam, USA), Cyclin D1 (1:1000, ab134175, Abcam, USA), Cyclin D3 (1:1000, ab28283, Abcam, USA), Cdc 42 (1:1000, ab187643, Abcam, USA), caspase-3 (1:1000, ab208161, Abcam, USA), and PARP (1:1000, ab32064, Abcam, USA).

Techniques: Mouse Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Experimental & Molecular Medicine

Article Title: Rab25 augments cancer cell invasiveness through a β1 integrin/EGFR/VEGF-A/Snail signaling axis and expression of fascin

doi: 10.1038/emm.2017.248

Figure Lengend Snippet: VEGF-A and VEGFR-1 are important for Snail expression. ( a ) Cells were transfected with indicated vectors. Quantitative RT-PCR (mean±s.d. * P

Article Snippet: Immunohistochemistry was performed with primary Rab25 (ab106175, 1:200, Abcam, Cambridge, MA, USA), VEGFR-1 (ab32152, 1:250, Abcam), Snail (ab180714, 1:100, Abcam) and fascin (ab126772, 1:250, Abcam) antibodies, as described previously.

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: Cell Death and Differentiation

Article Title: p73 is required for endothelial cell differentiation, migration and the formation of vascular networks regulating VEGF and TGFβ signaling

doi: 10.1038/cdd.2014.214

Figure Lengend Snippet: Lack of p73 affects TGF- β signaling in vivo . Quantification of VEGF-A expression and analysis of the TGF -β /ALK1/ID1 signaling pathway in P5 retinas from WT and p73KO mice. qRT-PCR analysis demonstrated a significant decrease in VEGF-A

Article Snippet: Western blotting was performed as previously described with the following primary antibodies: rabbit anti-pSmad1/5 (Ser463/465) 1 : 1000 (Cell Signaling, Danvers, MA, USA), rabbit anti Smad1 1 : 1000 (Cell Signaling), mouse anti-pERK 1 : 1000 (Santa Cruz Biotechnology), rabbit anti-ERK1 : 10 000 (Santa Cruz Biotechnology), rabbit anti-VEGF-A 1 : 1000 (Abcam) mouse anti DNp73 1 : 1000 (Imgenex, San Diego, CO, USA), and rabbit anti Nanog 1 : 1000 (Chemicon, Billerica, TX, USA) followed by the appropriate HRP-conjugated secondary antibodies (Pierce, Waltham, MA, USA).

Techniques: In Vivo, Expressing, Mouse Assay, Quantitative RT-PCR