primary antibodies against zeb2  (Novus Biologicals)

Journal: Clinical Cancer Research

Article Title: Overexpression of ZEB2 at the Invasion Front of Colorectal Cancer Is an Independent Prognostic Marker and Regulates Tumor Invasion In Vitro

doi: 10.1158/1078-0432.ccr-10-2816

Figure Lengend Snippet: Figure 1. Immunohistochemical analysis of ZEB2 in colorectal liver metastases (A–D) and primary colorectal cancer (E–H). Figures represent invasion front-specific overexpression of ZEB2 in paraffin- embedded specimens. Original magnification 200 (left) and corresponding areas (boxed areas) with lower magnification 80.

Article Snippet: Primary antibodies against ZEB2 (Rabbit polyclonal, Novus Biologicals, 1:200) and IgG-negative control (Mouse IgG, BD Pharmingen, 1:200) were incubated at 4 C overnight.

Techniques: Immunohistochemical staining, Over Expression

Journal: Clinical Cancer Research

Article Title: Overexpression of ZEB2 at the Invasion Front of Colorectal Cancer Is an Independent Prognostic Marker and Regulates Tumor Invasion In Vitro

doi: 10.1158/1078-0432.ccr-10-2816

Figure Lengend Snippet: Figure 2. DLD-1 cells were transfected with ZEB2 siRNA or scrambled control siRNA. Transfection efficiency was evaluated by qRT-PCR and reached more than 90% (A). DLD-1 cells were subjected to migration assays (B) and invasion assays (C). ZEB2 siRNA–transfected DLD-1 cells displayed an impaired migration capacity of 25% (P ¼ 0.018) and a reduced invasion capacity of 26% (P ¼ 0.001) compared with control. All assays were carried out 3 times in quadruplicates.

Article Snippet: Primary antibodies against ZEB2 (Rabbit polyclonal, Novus Biologicals, 1:200) and IgG-negative control (Mouse IgG, BD Pharmingen, 1:200) were incubated at 4 C overnight.

Techniques: Transfection, Control, Quantitative RT-PCR, Migration

Journal: Clinical Cancer Research

Article Title: Overexpression of ZEB2 at the Invasion Front of Colorectal Cancer Is an Independent Prognostic Marker and Regulates Tumor Invasion In Vitro

doi: 10.1158/1078-0432.ccr-10-2816

Figure Lengend Snippet: Figure 3. Kaplan–Meier curves display tumor-specific overall survival in patients with primary colorectal cancer (n ¼ 175). Solid line: negative/low expression of ZEB2; dashed line: high expression of ZEB2. A, overexpression of ZEB2 at the invasion front is significantly associated with a shortened tumor- specific survival (log-rank test, P 0.0001). B, overexpression of ZEB2 in the tumor center (Tc) correlated also with a shortened survival, but failed slightly to be statistically significant (log-rank test, P ¼ 0.06).

Article Snippet: Primary antibodies against ZEB2 (Rabbit polyclonal, Novus Biologicals, 1:200) and IgG-negative control (Mouse IgG, BD Pharmingen, 1:200) were incubated at 4 C overnight.

Techniques: Expressing, Over Expression

Journal: Reproductive Sciences

Article Title: Endometrial miR-200c is Altered During Transformation into Cancerous States and Targets the Expression of ZEBs , VEGFA , FLT1 , IKKβ , KLF9 , and FBLN5

doi: 10.1177/1933719112438448

Figure Lengend Snippet: Relative expression of miR-200c, ZEB1, and ZEB2, in endometrial biopsies (A) from mid-late proliferative phase (MLP), early- (ES), mid- (MS), and late (LS)-secretory phase of the menstrual cycle and endometrial tissues (B) from proliferative (Pro) and secretory (Sec) phases of the menstrual cycle, peri- and postmenopausal period (PPM), women exposed to GnRH agonist (GnRHa) and Depo-Provera (Depo), and grade I, II, and III endometrial cancer (C). The results are presented as mean ± standard error of the mean (SEM) and analyzed using analysis of variance (ANOVA) or nonparametric student t test (*P < .05 as compared to levels in mid-late proliferative phase [MLP] in Figure A, proliferative phase [Pro] in Figure B and peri- and postmenopausal period [PPM] in Figure C).

Article Snippet: Aliquots of 30 μg of total protein were subjected to Western blot analysis and transferred to polyvinylidene difluoride (PVDF) membrane, and then the membranes were probed with primary antibodies (dilution:1:200) generated against ZEB1, ZEB2, VEGFA, FLT1, FBLN5, TIMP2, IKKβ, and KLF9 (Cell Signaling Technology, Danvers, Massachusetts; Abcam Inc, San Francisco, California; Santa Cruz Biotechnology, Santa Cruz, California). α-tubulin (1:2000) was used for normalization and loading control.

Techniques: Expressing

Journal: Reproductive Sciences

Article Title: Endometrial miR-200c is Altered During Transformation into Cancerous States and Targets the Expression of ZEBs , VEGFA , FLT1 , IKKβ , KLF9 , and FBLN5

doi: 10.1177/1933719112438448

Figure Lengend Snippet: Relative expression of miR-200c, ZEB1, and ZEB2 in Ishikawa cells treated with 17-β Estradiol (E2) (A), progesterone (P4) (B), or medroxy progesterone acetate (MPA) (C) for 6 and 24 hours as compared to untreated control (Ctrl). The results are presented as mean ± standard error of the mean (SEM) and analyzed by analysis of variance (ANOVA) and nonparametric student t test (*P < .05 from their respective Ctrl).

Article Snippet: Aliquots of 30 μg of total protein were subjected to Western blot analysis and transferred to polyvinylidene difluoride (PVDF) membrane, and then the membranes were probed with primary antibodies (dilution:1:200) generated against ZEB1, ZEB2, VEGFA, FLT1, FBLN5, TIMP2, IKKβ, and KLF9 (Cell Signaling Technology, Danvers, Massachusetts; Abcam Inc, San Francisco, California; Santa Cruz Biotechnology, Santa Cruz, California). α-tubulin (1:2000) was used for normalization and loading control.

Techniques: Expressing, Control

Journal: Reproductive Sciences

Article Title: Endometrial miR-200c is Altered During Transformation into Cancerous States and Targets the Expression of ZEBs , VEGFA , FLT1 , IKKβ , KLF9 , and FBLN5

doi: 10.1177/1933719112438448

Figure Lengend Snippet: Relative expression of ZEB1 (A), ZEB2 (B), and CDH1 (C), in Ishikawa cells transfected with pre-miR-200c or pre-miR negative control (NC) determined by real-time polymerase chain reaction (PCR). The results are presented as mean ± standard error of the mean (SEM) and analyzed using nonparametric student t test with P values indicated by corresponding lines.

Article Snippet: Aliquots of 30 μg of total protein were subjected to Western blot analysis and transferred to polyvinylidene difluoride (PVDF) membrane, and then the membranes were probed with primary antibodies (dilution:1:200) generated against ZEB1, ZEB2, VEGFA, FLT1, FBLN5, TIMP2, IKKβ, and KLF9 (Cell Signaling Technology, Danvers, Massachusetts; Abcam Inc, San Francisco, California; Santa Cruz Biotechnology, Santa Cruz, California). α-tubulin (1:2000) was used for normalization and loading control.

Techniques: Expressing, Transfection, Negative Control, Real-time Polymerase Chain Reaction

Journal: Reproductive Sciences

Article Title: Endometrial miR-200c is Altered During Transformation into Cancerous States and Targets the Expression of ZEBs , VEGFA , FLT1 , IKKβ , KLF9 , and FBLN5

doi: 10.1177/1933719112438448

Figure Lengend Snippet: Western blot analysis of ZEB1, CDH1, VEGFA, IKKβ, KLF9, FLT1 and FBLN5 in Ishikawa cells transfected with pre-miR-200c and pre-miR negative control (NC) with α-Tubulin serving as loading control.

Article Snippet: Aliquots of 30 μg of total protein were subjected to Western blot analysis and transferred to polyvinylidene difluoride (PVDF) membrane, and then the membranes were probed with primary antibodies (dilution:1:200) generated against ZEB1, ZEB2, VEGFA, FLT1, FBLN5, TIMP2, IKKβ, and KLF9 (Cell Signaling Technology, Danvers, Massachusetts; Abcam Inc, San Francisco, California; Santa Cruz Biotechnology, Santa Cruz, California). α-tubulin (1:2000) was used for normalization and loading control.

Techniques: Western Blot, Transfection, Negative Control, Control

Journal: Reproductive Sciences

Article Title: Endometrial miR-200c is Altered During Transformation into Cancerous States and Targets the Expression of ZEBs , VEGFA , FLT1 , IKKβ , KLF9 , and FBLN5

doi: 10.1177/1933719112438448

Figure Lengend Snippet: Firefly luciferase assay with pZEX-MT01 or pGL3-Luc constructs carrying a 3′ untranslated region (3′ UTR) fragment of ZEB1 (A), ZEB2 (B), VEGFA (C), IKKβ (D), and KLF9 (E), respectively. Ishikawa cells were co-transfected with firefly luciferase reporters, Renilla luciferase transfection control plasmid (in case of IKKβ ), pre-miR-200c, or pre-miR negative control (NC). The ratio of firefly: Renilla was determined and reported as relative luciferase activity as compared to empty vector and analyzed using non-parametric student t-test with p values indicated by corresponding lines.

Article Snippet: Aliquots of 30 μg of total protein were subjected to Western blot analysis and transferred to polyvinylidene difluoride (PVDF) membrane, and then the membranes were probed with primary antibodies (dilution:1:200) generated against ZEB1, ZEB2, VEGFA, FLT1, FBLN5, TIMP2, IKKβ, and KLF9 (Cell Signaling Technology, Danvers, Massachusetts; Abcam Inc, San Francisco, California; Santa Cruz Biotechnology, Santa Cruz, California). α-tubulin (1:2000) was used for normalization and loading control.

Techniques: Luciferase, Construct, Transfection, Control, Plasmid Preparation, Negative Control, Activity Assay

Journal: Reproductive Sciences

Article Title: Endometrial miR-200c is Altered During Transformation into Cancerous States and Targets the Expression of ZEBs , VEGFA , FLT1 , IKKβ , KLF9 , and FBLN5

doi: 10.1177/1933719112438448

Figure Lengend Snippet: The miR-200c seed sequence and its complementary binding site on respective 3′ untranslated region (UTR) of its predicated target genes (represented in bold letters)

Article Snippet: Aliquots of 30 μg of total protein were subjected to Western blot analysis and transferred to polyvinylidene difluoride (PVDF) membrane, and then the membranes were probed with primary antibodies (dilution:1:200) generated against ZEB1, ZEB2, VEGFA, FLT1, FBLN5, TIMP2, IKKβ, and KLF9 (Cell Signaling Technology, Danvers, Massachusetts; Abcam Inc, San Francisco, California; Santa Cruz Biotechnology, Santa Cruz, California). α-tubulin (1:2000) was used for normalization and loading control.

Techniques: Sequencing, Binding Assay