rabbit anti human anti phosphorylated stathmin polyclonal antibody  (Bioss)

Journal: Oncology Letters

Article Title: Siva 1 inhibits proliferation, migration and invasion by phosphorylating Stathmin in ovarian cancer cells

doi: 10.3892/ol.2017.6307

Figure Lengend Snippet: Sequences of real-time PCR primers.

Article Snippet: Subsequent to blocking with 5% skim milk (YILI, Hohhot, Inner Mongolia, China) at room temperature for 1 h, diluted by TBS Tween (TBST), the PVDF membrane was incubated with the following antibodies at 4°C overnight: rabbit anti-human anti-Siva 1 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA, cat. no., sc-48767), rabbit anti-human anti-cleaved caspase-3 polyclonal antibody (dilution, 1:1,000; Abcam, Cambridge, UK, cat. no., ab2302), goat anti-human anti-cleaved caspase-9 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., cat. no., sc-22182), rabbit anti-human anti-Bcl-2-like protein 4 (Bax) polyclonal antibody (dilution, 1:400; Boster, Hubei, Wuhan, China, cat. no., BA0315), rabbit anti-human anti-Bcl-2 polyclonal antibody (dilution, 1:400; Boster, cat. no., BA0412), rabbit anti-human anti-Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, Beijing, China, cat. no., bs-1902R), rabbit anti-human anti-phosphorylated Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, cat. no., bs-3431R), rabbit anti-human anti-α-tubulin polyclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab178484) or mouse anti-human anti-acetyl-α-tubulin monoclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab24610).

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Oncology Letters

Article Title: Siva 1 inhibits proliferation, migration and invasion by phosphorylating Stathmin in ovarian cancer cells

doi: 10.3892/ol.2017.6307

Figure Lengend Snippet: Siva 1 enhances phosphorylation of Stathmin. (A) Siva 1 does not affect Stathmin mRNA level. (B and C) Overexpression of Siva 1 enhances phosphorylation of Stathmin and acetylation of α-tubulin, but did not markedly affect the expression level of Stathmin and α-tubulin detected by western blot. ***P<0.001 vs. pCMV group; ns, no significance.

Article Snippet: Subsequent to blocking with 5% skim milk (YILI, Hohhot, Inner Mongolia, China) at room temperature for 1 h, diluted by TBS Tween (TBST), the PVDF membrane was incubated with the following antibodies at 4°C overnight: rabbit anti-human anti-Siva 1 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA, cat. no., sc-48767), rabbit anti-human anti-cleaved caspase-3 polyclonal antibody (dilution, 1:1,000; Abcam, Cambridge, UK, cat. no., ab2302), goat anti-human anti-cleaved caspase-9 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., cat. no., sc-22182), rabbit anti-human anti-Bcl-2-like protein 4 (Bax) polyclonal antibody (dilution, 1:400; Boster, Hubei, Wuhan, China, cat. no., BA0315), rabbit anti-human anti-Bcl-2 polyclonal antibody (dilution, 1:400; Boster, cat. no., BA0412), rabbit anti-human anti-Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, Beijing, China, cat. no., bs-1902R), rabbit anti-human anti-phosphorylated Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, cat. no., bs-3431R), rabbit anti-human anti-α-tubulin polyclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab178484) or mouse anti-human anti-acetyl-α-tubulin monoclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab24610).

Techniques: Over Expression, Expressing, Western Blot

Journal: Gland Surgery

Article Title: The prognostic role of a phospho-Stathmin 1 signature in breast cancer treated with neoadjuvant chemotherapy

doi: 10.21037/gs-22-628

Figure Lengend Snippet: Relationship between STMN1 and its phosphorylation-related proteins and clinical outcomes for breast cancer patients. (A) Relationship between Kaplan-Meier analysis of DFS and STMN1 level by IHC (400×). (B) Relationship between Kaplan-Meier analysis of DFS and STMN1 phosphorylation at Ser16 levelby IHC (400×). (C) Relationship between Kaplan-Meier analysis of DFS and STMN1 phosphorylation at Ser25 levelby IHC (400×). (D) Relationship between Kaplan-Meier analysis of DFS and STMN1 phosphorylation at Ser38 levelby IHC (400×). (E) Relationship between Kaplan-Meier analysis of DFS and STMN1 phosphorylation at Ser63 levelby IHC (400×). (F) Relationship between Kaplan-Meier analysis of DFS and GRP78 level by IHC (400×). DFS, disease-free survival.

Article Snippet: Polyclonal rabbit anti-human STMN1 Ser38 antibody (Cell Signaling Technologies) was diluted to 1:100 and incubated with samples in a humid chamber at 4 °C overnight. respectively.

Techniques: Phospho-proteomics

Journal: Gland Surgery

Article Title: The prognostic role of a phospho-Stathmin 1 signature in breast cancer treated with neoadjuvant chemotherapy

doi: 10.21037/gs-22-628

Figure Lengend Snippet: Relationship between STMN1 phosphorylation and pathologic response. (A) Correlation between pathologic response and STMN1 expression. (B) Correlation between pathologic response and STMN1 phosphorylation at Ser-16. (C) Correlation between pathologic response and STMN1 phosphorylation at Ser-25. (D) Correlation between pathologic response and STMN1 phosphorylation at Ser-38. (E) Correlation between pathologic response and STMN1 phosphorylation at Ser-63. (F) Correlation between pathologic response and GRP78. pCR, pathological complete response.

Article Snippet: Polyclonal rabbit anti-human STMN1 Ser38 antibody (Cell Signaling Technologies) was diluted to 1:100 and incubated with samples in a humid chamber at 4 °C overnight. respectively.

Techniques: Phospho-proteomics, Expressing

Journal: Gland Surgery

Article Title: The prognostic role of a phospho-Stathmin 1 signature in breast cancer treated with neoadjuvant chemotherapy

doi: 10.21037/gs-22-628

Figure Lengend Snippet: p-STMN1/GRP78 model powerfully predicts DFS for patients treated with NACT. (A) Kaplan-Meier analysis of DFS in breast cancer patients from high and low-risk groups classified by the p-STMN1/GRP78 model. (B) ROC curves to test the prognostic accuracy of the p-STMN1/GRP78 model. DFS, disease-free survival; ROC, receiver operating characteristic; AUC, area under the receiver operating characteristic curve.

Article Snippet: Polyclonal rabbit anti-human STMN1 Ser38 antibody (Cell Signaling Technologies) was diluted to 1:100 and incubated with samples in a humid chamber at 4 °C overnight. respectively.

Techniques:

Journal: Oncology Letters

Article Title: Siva 1 inhibits proliferation, migration and invasion by phosphorylating Stathmin in ovarian cancer cells

doi: 10.3892/ol.2017.6307

Figure Lengend Snippet: Sequences of real-time PCR primers.

Article Snippet: Subsequent to blocking with 5% skim milk (YILI, Hohhot, Inner Mongolia, China) at room temperature for 1 h, diluted by TBS Tween (TBST), the PVDF membrane was incubated with the following antibodies at 4°C overnight: rabbit anti-human anti-Siva 1 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA, cat. no., sc-48767), rabbit anti-human anti-cleaved caspase-3 polyclonal antibody (dilution, 1:1,000; Abcam, Cambridge, UK, cat. no., ab2302), goat anti-human anti-cleaved caspase-9 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., cat. no., sc-22182), rabbit anti-human anti-Bcl-2-like protein 4 (Bax) polyclonal antibody (dilution, 1:400; Boster, Hubei, Wuhan, China, cat. no., BA0315), rabbit anti-human anti-Bcl-2 polyclonal antibody (dilution, 1:400; Boster, cat. no., BA0412), rabbit anti-human anti-Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, Beijing, China, cat. no., bs-1902R), rabbit anti-human anti-phosphorylated Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, cat. no., bs-3431R), rabbit anti-human anti-α-tubulin polyclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab178484) or mouse anti-human anti-acetyl-α-tubulin monoclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab24610).

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Oncology Letters

Article Title: Siva 1 inhibits proliferation, migration and invasion by phosphorylating Stathmin in ovarian cancer cells

doi: 10.3892/ol.2017.6307

Figure Lengend Snippet: Siva 1 enhances phosphorylation of Stathmin. (A) Siva 1 does not affect Stathmin mRNA level. (B and C) Overexpression of Siva 1 enhances phosphorylation of Stathmin and acetylation of α-tubulin, but did not markedly affect the expression level of Stathmin and α-tubulin detected by western blot. ***P<0.001 vs. pCMV group; ns, no significance.

Article Snippet: Subsequent to blocking with 5% skim milk (YILI, Hohhot, Inner Mongolia, China) at room temperature for 1 h, diluted by TBS Tween (TBST), the PVDF membrane was incubated with the following antibodies at 4°C overnight: rabbit anti-human anti-Siva 1 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., Dallas, TX, USA, cat. no., sc-48767), rabbit anti-human anti-cleaved caspase-3 polyclonal antibody (dilution, 1:1,000; Abcam, Cambridge, UK, cat. no., ab2302), goat anti-human anti-cleaved caspase-9 polyclonal antibody (dilution, 1:200; Santa Cruz Biotechnology, Inc., cat. no., sc-22182), rabbit anti-human anti-Bcl-2-like protein 4 (Bax) polyclonal antibody (dilution, 1:400; Boster, Hubei, Wuhan, China, cat. no., BA0315), rabbit anti-human anti-Bcl-2 polyclonal antibody (dilution, 1:400; Boster, cat. no., BA0412), rabbit anti-human anti-Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, Beijing, China, cat. no., bs-1902R), rabbit anti-human anti-phosphorylated Stathmin polyclonal antibody (dilution, 1:500; Bioss Antibodies, cat. no., bs-3431R), rabbit anti-human anti-α-tubulin polyclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab178484) or mouse anti-human anti-acetyl-α-tubulin monoclonal antibody (dilution, 1:1,000; Abcam, cat. no., ab24610).

Techniques: Over Expression, Expressing, Western Blot

Journal: Scientific Reports

Article Title: Downregulation of stathmin 1 in human gallbladder carcinoma inhibits tumor growth in vitro and in vivo

doi: 10.1038/srep28833

Figure Lengend Snippet: Representative images of STMN1 immunostaining in adjacent normal tissues ( a,b ) and different stages of GBC tissues (GBC tissue of Grade II ( c,d ); GBC tissue of Grade II~III, III ( e,f )). Twelve pairs of GBC and pericarcinomatous tissues were sectioned and subjected to immunohistochemistry, as described in Methods section (Origianl magnification is 200×). Red arrows indicated STMN1 positive signal. The expression levels of STMN1 between GBC and adjacent tissue were shown in the scatter plot ( p = 0.0003, significance was determined using Student’s t -test) ( g ). The STMN1 expression level was scored by semi-quantitative scoring system as described in Methods section. The protein level of STMN1 was detected in two GBC cell lines (GBC-SD, and SGC-996) ( h ), with GBC and pericarcinomatous tissue were used as positive and negative controls, respectively. GAPDH was used as a control for protein loading. NT: adjacent normal tissues; T: tumor tissue. GBC: Gallbladder carcinoma.

Article Snippet: The slides were then incubated with rabbit anti-human STMN1 polyclonal antibodies (1:600; Cell Signaling Technology, MA, USA) overnight at 4 °C, followed by incubation with a secondary antibody (UltraSensitive SP; Fuzhou Maixin Biotech.

Techniques: Immunostaining, Immunohistochemistry, Expressing, Control

Journal: Scientific Reports

Article Title: Downregulation of stathmin 1 in human gallbladder carcinoma inhibits tumor growth in vitro and in vivo

doi: 10.1038/srep28833

Figure Lengend Snippet: SGC-996 and GBC-SD cells were transfected with vector control (NC) or shSTMN1-1. 72 h later, the protein of each cell line was extracted. The expression of STMN1 in the two cell lines was detected by WB ( a ) and qRT-PCR ( b ), respectively. Then, shSTMN1-1 was selected randomly for further research. The growth curve was tested by MTS assay ( c ). Migration ( d ) and invasion ( e ) assays of SGC-996 and GBC-SD cells were performed 24 h after incubated. Results were shown as means ± SEM from three independent experiments. NS: Non-significant; NC: Negative control. * p < 0.05; ** p < 0.01. Significance was determined using Student’s t -test.

Article Snippet: The slides were then incubated with rabbit anti-human STMN1 polyclonal antibodies (1:600; Cell Signaling Technology, MA, USA) overnight at 4 °C, followed by incubation with a secondary antibody (UltraSensitive SP; Fuzhou Maixin Biotech.

Techniques: Transfection, Plasmid Preparation, Control, Expressing, Quantitative RT-PCR, MTS Assay, Migration, Incubation, Negative Control

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: A. Representative IHC staining of high and low expression of STMN1 in the large (400×) and small images (100×). B. Representative IHC staining of high and low expression of multiple phosphor-sites (Ser-16, Ser-25, Ser38, Ser63) in the large (400×) and small images (100×). C. Kaplan-Meier analysis of DFS in the training set. D. Kaplan-Meier analysis of DFS in the validation set.

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Immunohistochemistry, Expressing, Biomarker Discovery

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: Univariate association of the STMN1-E/P model, clinicopathological characteristics, and single phospho-sites status with disease-free survival

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Biomarker Discovery

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: Data are shown as AUC (95% CI) or hazard ratios (95% CI). ROC = receiver operator characteristic. AUC = area under the curve. A. Comparisons of the prognostic accuracy by the STMN1-E/P model and TNM stage in the training set. B. DFS of patients with high- or low-risk scores in the training set. C. Comparisons of the prognostic accuracy by the STMN1-E/P model and TNM stage in the validation set. D. DFS of patients with high- or low-risk scores in the validation set. P values were calculated using the log-rank test.

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Biomarker Discovery

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: A. Comparisons of the prognostic accuracy by the STMN1-E/P/C model, STMN1-E/P model and TNM stage in the training set. B. DFS of patients with high- or low-risk scores according to the STMN1-E/P/C model in the training set. C. Comparisons of the prognostic accuracy by the STMN1-E/P/C model, STMN1-E/P model and TNM stage in the validation set. D. DFS of patients with high- or low-risk scores according to the STMN1-E/P/C model in the validation set. P values were calculated using the log-rank test. E. DFS of patients with luminal breast cancer. F. DFS of patients with HER2/neu subtype breast cancer. G. DFS of patients with TNBC subtype breast cancer.

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Biomarker Discovery

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: A. Representative IHC staining of high and low expression of STMN1 in the large (400×) and small images (100×). B. Representative IHC staining of high and low expression of multiple phosphor-sites (Ser-16, Ser-25, Ser38, Ser63) in the large (400×) and small images (100×). C. Kaplan-Meier analysis of DFS in the training set. D. Kaplan-Meier analysis of DFS in the validation set.

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Immunohistochemistry, Expressing, Biomarker Discovery

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: Univariate association of the STMN1-E/P model, clinicopathological characteristics, and single phospho-sites status with disease-free survival

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Biomarker Discovery

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: Data are shown as AUC (95% CI) or hazard ratios (95% CI). ROC = receiver operator characteristic. AUC = area under the curve. A. Comparisons of the prognostic accuracy by the STMN1-E/P model and TNM stage in the training set. B. DFS of patients with high- or low-risk scores in the training set. C. Comparisons of the prognostic accuracy by the STMN1-E/P model and TNM stage in the validation set. D. DFS of patients with high- or low-risk scores in the validation set. P values were calculated using the log-rank test.

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Biomarker Discovery

Journal: Oncotarget

Article Title: Stathmin and phospho-stathmin protein signature is associated with survival outcomes of breast cancer patients

doi:

Figure Lengend Snippet: A. Comparisons of the prognostic accuracy by the STMN1-E/P/C model, STMN1-E/P model and TNM stage in the training set. B. DFS of patients with high- or low-risk scores according to the STMN1-E/P/C model in the training set. C. Comparisons of the prognostic accuracy by the STMN1-E/P/C model, STMN1-E/P model and TNM stage in the validation set. D. DFS of patients with high- or low-risk scores according to the STMN1-E/P/C model in the validation set. P values were calculated using the log-rank test. E. DFS of patients with luminal breast cancer. F. DFS of patients with HER2/neu subtype breast cancer. G. DFS of patients with TNBC subtype breast cancer.

Article Snippet: For STMN1 and Ser38, TMAs were blocked with 10% normal goat serum for 1 h at room temperature (RT) and incubated in a humid chamber at 4°C overnight with polyclonal rabbit anti-human STMN1 antibody (Proteintech) or polyclonal rabbit anti-human STMN1 Ser38 (Cell Signaling Technologies) antibody diluted to 1:400 or 1:100, respectively.

Techniques: Biomarker Discovery

Journal: Oncoscience

Article Title: P18/Stathmin1 is regulated by miR-31 in ovarian cancer in response to taxane

doi:

Figure Lengend Snippet: Differentia expression of STMN1 and miR-31 in ovarian cancer tissue samples

Article Snippet: Rabbit anti-human stathmin1 (STMN1) polyclonal antibody (Cell Signaling Technology, #3352, Danvers, MA, USA) was used for immunohistochemistry.

Techniques: Expressing

Journal: Oncoscience

Article Title: P18/Stathmin1 is regulated by miR-31 in ovarian cancer in response to taxane

doi:

Figure Lengend Snippet: A. Representative results of IHC with high expression (+++; a) or very weak expression (+; b) are shown as photographs. B. Conclusive diagram shows the relationship between the expression of STMN1and chemoresponse In the ovarian cancer tissues examined. The high STMAN expression is significantly correlated with chemoresistance. From 24 ovarian cancer tissues samples, 14 tissue samples showed positive STMN1 expression. Of them, 11 samples (78.5%) were chemoresistant while 3 samples (21.5%) were chemoresponsive. On the other hand, ten samples showed negative or very week STMN1 expression. C. Western blotting analysis of STMN1 expression in four cell lines. Chemoresistant KF-TX cells express more STMN1 compared with parental KF cells. The blot shows that STMN1 expression is associated with IC50 to TX. SKOV-3, lowest IC50, shows the lowest expression level of STMN1.

Article Snippet: Rabbit anti-human stathmin1 (STMN1) polyclonal antibody (Cell Signaling Technology, #3352, Danvers, MA, USA) was used for immunohistochemistry.

Techniques: Expressing, Western Blot

Journal: Oncoscience

Article Title: P18/Stathmin1 is regulated by miR-31 in ovarian cancer in response to taxane

doi:

Figure Lengend Snippet: A. A representative western blot shows the amendment of STMN1-specific but not control siRNA in KF-TX cells. Tubulin immunoblot was used as a loading control. The experiment was repeated three independent times for reproducibility. B. KF-TX cells were transfected with STMN1 siRNA or control siRNA (100 nM) twice. One day after last transfection, equal cell numbers were subcultured for further 24h, and then treated with TX for three days. The dose response curves are shown as Hill equations while each data point represents mean of three experiments; bars denote S.E;, C. Annexin V staining after TX in STMN1 KD cells compared with control. KF-TX were treated after SMN1 KD as well as in B. and then stained by annexin V and acquired by FACS analyzer. The cytograph shows a significant enhancement of TX-induced apoptosis as indicated by annexin stained cells.

Article Snippet: Rabbit anti-human stathmin1 (STMN1) polyclonal antibody (Cell Signaling Technology, #3352, Danvers, MA, USA) was used for immunohistochemistry.

Techniques: Western Blot, Control, Transfection, Staining

Journal: Oncoscience

Article Title: P18/Stathmin1 is regulated by miR-31 in ovarian cancer in response to taxane

doi:

Figure Lengend Snippet: A. Representative RT–PCR show the expression levels of miR-31 in cell clones named as clones (#1) and (#2) are high and middle, respectively B. STMN1 expression responses to miR-31 expression. Immunobloting analysis of STMN1 in the miR-31 overexpressing clones and control ones. The KF-TX-miR-31 show less expression level of STMN1 compared with control (left panel). Representative RT-PCR in one stable clone (#1) and its control shows that the STMN1 transcript level decreased compared with the control clone (right panel)

Article Snippet: Rabbit anti-human stathmin1 (STMN1) polyclonal antibody (Cell Signaling Technology, #3352, Danvers, MA, USA) was used for immunohistochemistry.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Clone Assay, Western Blot, Control, Stable Transfection

Journal: Oncoscience

Article Title: P18/Stathmin1 is regulated by miR-31 in ovarian cancer in response to taxane

doi:

Figure Lengend Snippet: A. KF-TX cells show a differential expression profile of STMN1 in response to TX (different doses for 48 h) in miR-31 overexpressing cells compared to control. The KF-TX-miR-31 show less expression level of STMN1 gradually increase at 50nm treatment point and again decrease at 200nm. The Control (GFP) clone show relatively high expression of STMN1 which significantly increased and did not show significant reduction even at 200nm. B. miR-31 restores response to TX in KF-TX cells. Viability assay shows the differential response to TX in KF-TX-miR-31 (clones #1 and #2) and representative control (GFP only) clone (#1) for three days. The relative viability show that the clones overexpressing miR-31 are significantly sensitive to TX compared with the control. C. A representative FACS analysis shows the different apoptotic outcome of KF-TX cells clones overexpressing miR-31 (#1) and representative control clone (#1) after treatment with 250nM TX for the indicated time. D. Phase contrast of two different clones; control #2 and miR-31 #2 after 3-days of TX treatment at different doses for three days. E. Effect of miR-31 inhibitor on STMN1 expression. Immunobloting analysis of STMN1 in the parental cells, KF, transfected with the anti-miR-31 shows the relative upregulation of STMN1 compared with the control inhibitor (upper panel). Effect of anti-miR-31 on chemosensitivity to TX analyzed by viability assay in the parental KF cells. Anti-miR-31 confers chemoresistance in KF cells.

Article Snippet: Rabbit anti-human stathmin1 (STMN1) polyclonal antibody (Cell Signaling Technology, #3352, Danvers, MA, USA) was used for immunohistochemistry.

Techniques: Quantitative Proteomics, Control, Expressing, Viability Assay, Clone Assay, Western Blot, Transfection

Journal: Oncoscience

Article Title: P18/Stathmin1 is regulated by miR-31 in ovarian cancer in response to taxane

doi:

Figure Lengend Snippet: A. Acetylated α-tubulin in the far immunoblot of α-tubulin immunoprecipitant from cell lysates isolated from either control/GFP or two stable clones overexpressing miR-31(#1 and #2). Acetylated tubulin is greater in KF-TX cells which overexpress miR-31, with depleted STMN1, compared with control transfectant which show higher STMN1. B. Immunocytochemical staining shows tubulin in KF-TX cells overexpressing miR-31 and their control (GFP). Representative images of a miR-31 clone (#1) and a control clone (#1) were fixed and stained with antibodies against α-tubulin. miR-31 overexpressing cells show more polymerized and bundled tubulin nearby the margins of the cells compared with normal tubulin distribution in the control as an evidence for STMN1 depletion in miR-31 clone compared with control one. miR-31 overexpression slows interphase in KF-TX cells. C. Phase contrast image obtained from time laps microscopic study (additional files; 1, 2). The series for a cell exiting mitosis. Plot summarizes data from >10 cells and three independent experiments per condition. STMN1 depletion increased interphase duration by approximately 210 min (**, P <0.05), compared with cells progressed through mitosis with normal kinetics. D. Immunoflorcent images show the distribution of the tubulin in the representative clones (KF-TX-miR-31; clone#1 and control-GFP #1). The miR-31 overexpressing cells showed bundled tubulin nearby the margimes of the cytoplasm E. Luciferase activity after transfection of the indicated 3′-UTR-driven reporter constructs. Reporter plasmid containing 3′-UTR region of STMN1 with binding site of miR-31 or with the binding site deleted, *P< 0.05.

Article Snippet: Rabbit anti-human stathmin1 (STMN1) polyclonal antibody (Cell Signaling Technology, #3352, Danvers, MA, USA) was used for immunohistochemistry.

Techniques: Western Blot, Isolation, Control, Clone Assay, Transfection, Staining, Over Expression, Luciferase, Activity Assay, Construct, Plasmid Preparation, Binding Assay

Journal: Oncology reports

Article Title: Stathmin expression in glioma-derived microvascular endothelial cells: a novel therapeutic target.

doi: 10.3892/or.2011.1525

Figure Lengend Snippet: Figure 1. Immunohistochemical staining for FVIII-Rag in cultured GDMEC (DAB staining appears brown-colored; original magnification, x400). (B-D) Analysis of stathmin expression in human glioma by immunohistochemistry. Stathmin was detected mainly in the cytoplasm of VEC from microvessels and in glial cells (original magnification, x400; counterstaining with H&E). MV, microvessel; EC, endothelial cell. (B) Expression of stathmin in normal brain. (C) Expression of stathmin in low-grade glioma (WHO grade I-II). (D) Expression of stathmin in high grade glioma (WHO grade III-IV). (E) Western blot analysis of stathmin protein expression in GDMEC and NDMEC, lanes: 1, GDMEC from WHO I-II tumors; 2, GDMEC from grade III-IV tumors; 3, NDMEC. (F) RT-PCR analysis of stathmin mRNA in GDMEC and NDMEC, lanes: 1, GDMEC from WHO I-II tumors; 2, GDMEC from WHO III-IV tumors; 3, NDMEC.

Article Snippet: A rabbit anti-human stathmin polyclonal primary antibody (CST, USA) was applied in overnight incubation.

Techniques: Immunohistochemical staining, Staining, Cell Culture, Expressing, Immunohistochemistry, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Oncology reports

Article Title: Stathmin expression in glioma-derived microvascular endothelial cells: a novel therapeutic target.

doi: 10.3892/or.2011.1525

Figure Lengend Snippet: Figure 2. (A) Western blot analysis on stathmin expression of siRNA transfection. (B) RT-PCR analysis on stathmin transcription of siRNA transfection. (C) Proliferation of GDMEC in the siRNA group is suppressed, the difference between the siRNA and control groups is statistically significant (*P<0.05). (D and E) The apoptosis rate of siRNA-transfected GDMEC is significantly higher than the control group (*P<0.05). (F) Western blot demonstrates that expression of apoptosis associated proteins. Bcl-X, Bcl-2 and pro-caspase-3 were down-regulated while Bax was up-regulated in the siRNA-transfected group.

Article Snippet: A rabbit anti-human stathmin polyclonal primary antibody (CST, USA) was applied in overnight incubation.

Techniques: Western Blot, Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Control

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Immunohistochemical staining of stathmin1 in oral and gastric cancer sections. Anti-stathmin1 antibody was used for immunohistochemical staining in human oral and gastric cancer tissues as described in ‘Materials and Methods’ section. The invading oral cancer cells were positive for stathmin1 protein in the cytoplasm. The normal gastric mucosal cells were either negative or very weakly positive for stathmin1 protein in the cytoplasm. The invading gastric cancer cells were positive for stathmin1 protein. Scale bar, 200 μ m.

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Immunohistochemical staining, Staining

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Correlation between the expression of stathmin1 and clinical classification in gastric cancer

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Expressing

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Correlationship between the stathmin1 expression level and clinicopathological characteristics. Data of stathmin-positive patients with diffuse-type gastric cancer are presented. ( A ) The state of stathmin1 protein expression in patients with lymph node metastasis in diffuse type gastric cancer ( n =17) and in patients without metastasis ( n =7). Stathmin1 protein expression in patients with lymph node metastasis is significantly higher than in patients without lymph node metastasis ( * P <0.05, Mann–Whitney U -test). ( B ) The state of stathmin1 protein expression in patients with early stages (I and II) of diffuse type gastric cancer ( n =9) and patients with advanced stages (III and IV) of diffuse-type gastric cancer ( n =15). Stathmin1 protein expression in advanced stages is significantly higher than that in early stages ( ** P <0.01, Mann–Whitney U -test). ( C ) The state of stathmin1 protein expression in patients with vascular invasion in diffuse-type gastric cancer ( n =14) and in patients without vascular invasion ( n =10). Stathmin1 expression in patients with vascular invasion is significantly higher than in patients without vascular invasion ( ** P <0.01, Mann–Whitney U -test).

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Expressing, MANN-WHITNEY

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Kaplan–Meier curves for recurrence-free survival according to stathmin1 overexpression in patients with gastric cancer (( A ) diffuse type+intestinal type, P <0.05; ( B ) diffuse type only, P <0.01). When stathmin1 overexpression was observed, the recurrence-free survival rate decreased significantly, especially in patients with diffuse-type gastric cancer.

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Over Expression

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Univariate and multivariate analysis of prognostic factors in the diffuse type of gastric cancer patients for recurrence-free survival

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Significance Assay, Expressing

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Stathmin1 was specifically downregulated by stathmin1 siRNA. Cells were transfected with scrambled siRNA or stathmin1 siRNA. ( A ) Two days later, SNU638 and SNU16 cells were collected and stathmin1 protein levels were detected by western blotting. β -actin expression was monitored for normalisation. ( B ) SNU638 cells were collected and stathmin1 mRNA levels were examined by real-time PCR. Values are expressed as the percentage of control, which was defined as 100% ( n =6). Results were analysed by one-way ANOVA, followed by Tukey's multiple comparison test. * P <0.01 vs control. mRNA, messenger RNA; SCR, scrambled; siRNA, small interfering RNA.

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Transfection, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Control, Comparison, Small Interfering RNA

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Effect of stathmin1 silencing on proliferation of gastric cancer cells. SNU638 ( A ) or SNU16 ( B ) cells were transfected with SCR siRNA or stathmin1 ( STMN1 ) siRNA. After 5 days of incubation, cell proliferation was evaluated in WST assay. Data are expressed as percentage change (means±s.d.) compared with controls and represent four independent experiments. ( * P <0.05, ** P <0.01 vs SCR siRNA, one-way ANOVA followed by Tukey's multiple comparison). ANOVA, one-way analysis of variance; SCR, scrambled; siRNA, small interfering RNA.

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Transfection, Incubation, WST Assay, Comparison, Small Interfering RNA

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Effect of stathmin1 silencing on the migration and invasion of gastric cancer cells. ( A ) Cell migration was evaluated in the Boyden migration assay two days after SNU638 cells were transfected with scrambled (SCR) small interfering RNA (siRNA) or stathmin1 siRNA. ( B ) Cell invasion was evaluated in the Matrigel invasion assay as described in the ‘Materials and Methods’ section. Data are expressed as percentage change (means±s.d.) compared with controls and represent four independent experiments. ( * P <0.01 vs SCR siRNA, one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison). Representative microscopic images were presented in the upper panel of each assay graph. ANOVA, one-way analysis of variance; SCR, scrambled; siRNA, small interfering RNA.

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: Migration, Transfection, Small Interfering RNA, Invasion Assay, Comparison

Journal: British Journal of Cancer

Article Title: Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells

doi: 10.1038/sj.bjc.6605537

Figure Lengend Snippet: Effect of stathmin1 silencing on in vivo tumour growth. ( A ) SNU638 cells were transfected with scrambled (SCR) small interfering RNA (siRNA) or stathmin1 ( STMN1 ) siRNA, and nude mice were inoculated subcutaneously with 2 × 10 6 cells at two sites per mouse. The tumour mass (xenograft) volume was measured every week from week 3 to week 7. Data are expressed as the means±s.d. and represent four independent experiments. ( * P <0.05, ** P <0.01 vs SCR siRNA, one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison) ( B ) The tumour masses (xenograft) were dissected out 7 weeks later and presented. ANOVA, one-way analysis of variance; SCR, scrambled; siRNA, small interfering RNA.

Article Snippet: Immunohistochemical staining with rabbit anti-human stathmin1 polyclonal antibody (Cell Signaling Technology, Danvers, MA, USA, 1 : 50) and with rabbit anti-Ki-67 polyclonal antibody (Novocastra, Newcastle, UK, 1 : 200) was performed on 4- μ m sections of paraffin-embedded specimens.

Techniques: In Vivo, Transfection, Small Interfering RNA, Comparison