rabbit antihuman foxm1  (Cell Signaling Technology Inc)

Journal: Journal of the American Heart Association

Article Title: Wnt Site Signaling Inhibitor Secreted Frizzled‐Related Protein 3 Protects Mitral Valve Endothelium From Myocardial Infarction–Induced Endothelial‐to‐Mesenchymal Transition

doi: 10.1161/jaha.121.023695

Figure Lengend Snippet: Figure 7. FOXM1 contributes to EndMT and can be suppressed by sFRP3. A, Mitral VECs were treated with sham, post-MI plasma, and post-MI plasma with Siomycin A (1 µmol/L) for 24 hours before immunofluorescence staining using FOXM1 antibody (green; scale bar: 20 µm). B, Percentage of the cells positive for nuclear FOXM1 per total nuclei from 4 independent immunofluorescence assays were graphed. Statistical analysis was conducted using 1-way ANOVA with Tukey’s multiple comparisons test. C, Mitral VECs were treated with MI plasma (n=6) with and without Siomycin A (1 µmol/L) for 24 hours before qPCR analysis. Mean±SD from 3 independent assays were graphed. P values were calculated using the Wilcoxon signed-rank test and were significant (P<0.05) for all tested genes. D, Mitral VECs were treated with MI plasma (n=6) supplemented with sFRP3 (250 ng/mL) for 24 hours before qPCR analysis. Mean±SD from 3 independent assays were graphed. P value was calculated using nonparametric Mann- Whitney test. E, FOXM1 staining in mitral VECs following exposure to post-MI plasma and post-MI plasma supplemented with 250 ng/mL of sFRP3 for 24 hours. DAPI was used to stain nuclei (scale bar: 50 µm). F, Mean±SD of % cells positive for nuclear FOXM1 per total nuclei from 5 independent immunofluorescence assays with 2 individual post-MI plasma were graphed. P values were calculated using Mann-Whitney test. EndMT indicates endothelial-to-mesenchymal transition; FC, fold changes; FOXM1, forkhead box M1; MI, myocardial infarction; NS, not significant; qPCR, quantitative polymerase chain reaction; sFRP3, secreted frizzled-related protein 3; SioA, Siomycin A; and VEC, valve endothelial cell.

Article Snippet: Sections were then incubated with rabbit antihuman FOXM1 polyclonal antibody (Novus Biologicals, Littleton, CO; No. NBP1- 30961) diluted 1:300 or anti- Ki67 (Abcam; No. 15580) diluted 1:50 in 5% normal horse serum (Vector Laboratories, Burlingame, CA; No. S- 2000) for 90 minutes at room temperature, followed by 45 minutes incubation with antirabbit secondary antibody (LSAB Kit; Dako; No. K0675).

Techniques: Clinical Proteomics, Immunofluorescence, Staining, MANN-WHITNEY, Real-time Polymerase Chain Reaction

Journal: Journal of the American Heart Association

Article Title: Wnt Site Signaling Inhibitor Secreted Frizzled‐Related Protein 3 Protects Mitral Valve Endothelium From Myocardial Infarction–Induced Endothelial‐to‐Mesenchymal Transition

doi: 10.1161/jaha.121.023695

Figure Lengend Snippet: Figure 8. Proposed model of EndMT induction in MV by post-MI plasma. sFRP3-deficient post-MI plasma releases the brake on Wnt signaling, increases FOXM1 transcriptional activity, nuclear localization of Slug, which initiates EndMT within days after MI. EndMT indicates endothelial-to-mesenchymal transition; FOXM1, forkhead box M1; MI, myocardial infarction; MV, mitral valve; NS, not significant; sFRP3, secreted frizzled-related protein 3; TGFβ, transforming growth factor beta; VE, vascular endothelial; and Wnt, wingless-related integration site.

Article Snippet: Sections were then incubated with rabbit antihuman FOXM1 polyclonal antibody (Novus Biologicals, Littleton, CO; No. NBP1- 30961) diluted 1:300 or anti- Ki67 (Abcam; No. 15580) diluted 1:50 in 5% normal horse serum (Vector Laboratories, Burlingame, CA; No. S- 2000) for 90 minutes at room temperature, followed by 45 minutes incubation with antirabbit secondary antibody (LSAB Kit; Dako; No. K0675).

Techniques: Clinical Proteomics, Activity Assay

Journal: Journal of Breast Cancer

Article Title: High MicroRNA-370 Expression Correlates with Tumor Progression and Poor Prognosis in Breast Cancer

doi: 10.4048/jbc.2015.18.4.323

Figure Lengend Snippet: Representative microphotographs of immunohistochemical staining of FOX proteins in breast cancer (×400). The tumor cells shows positive nuclear staining. (A) FOXO1 (intensity score 2), (B) FOXO3a (intensity score 3), and (C) FOXM1 (intensity score 3).

Article Snippet: The primary antibodies were rabbit antihuman FOXM1 (sc-502; Santa Cruz Biotechnology, Santa Cruz, USA), FOXO1 (#2880; Cell Signaling, Danvers, USA) and FOXO3a (sc-11351; Santa Cruz Biotechnology) polyclonal antibodies used at a 1:100 dilution.

Techniques: Immunohistochemical staining, Staining

Journal: Journal of Breast Cancer

Article Title: High MicroRNA-370 Expression Correlates with Tumor Progression and Poor Prognosis in Breast Cancer

doi: 10.4048/jbc.2015.18.4.323

Figure Lengend Snippet: Association between miR-370 expression and FOX protein expression in breast cancers

Article Snippet: The primary antibodies were rabbit antihuman FOXM1 (sc-502; Santa Cruz Biotechnology, Santa Cruz, USA), FOXO1 (#2880; Cell Signaling, Danvers, USA) and FOXO3a (sc-11351; Santa Cruz Biotechnology) polyclonal antibodies used at a 1:100 dilution.

Techniques: Expressing

Journal: CNS Neuroscience & Therapeutics

Article Title: Curcumin Suppresses Malignant Glioma Cells Growth and Induces Apoptosis by Inhibition of SHH / GLI 1 Signaling Pathway in Vitro and Vivo

doi: 10.1111/cns.12163

Figure Lengend Snippet: Curcumin inhibited SHH/GLI1 signaling and its downstream targets oncoproteins in time‐ and dose‐dependent manner. (A) Real‐time everse transcription‐PCR analysis of total RNA from T98G cells following 40 uM of curcumin treatment for different time points (0, 8, 16, 24 h) revealed decreasing expression of Shh, Smo, and GLI1 on mRNA level. (B) Western blot assay showed curcumin treatment led to decreased levels of Shh, GLI1, and GLI1‐regulated target (CyclinD1, Bcl‐2, Foxm1) proteins in time‐dependent manner in T98G cells following 40 uM agent exposure for 24 h. (C) U87 and T98G cells were plated and grown for 24 h and then treated with the solvent DMSO (control) or with different (indicated) doses of curcumin dissolved in DMSO for 24 h before harvesting. Western blot assay shows curcumin inhibits core components of the SHH/GLI1 signaling (SHH, GLI1) and its target oncoproteins (CyclinD1, Bcl‐2, Foxm1) in dose‐dependent manner. Data are shown as mean ± SEM for the three replicates. Statistical significance levels are indicated as: *P < 0.05; **P < 0.01 and ***P < 0.001.

Article Snippet: The membrane was blocked in 5% milk–TBST solution and incubated separately with rabbit antihuman Shh (1:500, Bioss Inc, USA), rabbit antihuman GLI1 (1:1000, Cell Signaling Technology, USA), mouse antihuman CyclinD1 antibody (1:300, Santa Cruz, USA),rabbit antihuman Bcl‐2(1:500, Santa Cruz, USA), rabbit antihuman Foxm1(1:1000, Cell Signaling Technology, USA), rabbit antihuman Bax (1:500, Santa Cruz, USA), mouse antihuman pro‐caspase‐9 (1:500, Santa Cruz, USA), rabbit antihuman pro‐caspase‐3 (1:500, Santa Cruz, USA), or mouse antihuman GAPDH antibody (1:1000, Sigma, USA).

Techniques: Expressing, Western Blot