pstat5 antibody Search Results


anti-phospho-stat5 (p-stat5) alexa 488  (Becton Dickinson)
90
Becton Dickinson anti-phospho-stat5 (p-stat5) alexa 488
Flow cytometric gating strategy adopted to identify CD33+/CD34+ precursor cells. A representative JMML patient is showed. Mononuclear cells were initially gated to exclude debris and residual granulocytes by physical parameters ( a ); all myeloid cells were selected by their reactivity to anti-CD33 antibody ( b ); myeloid precursors were then identified as CD33+/CD34+ double positive cells ( c ). CD33+/CD34+ cells were further checked for their negativity to anti-CD14 antibody ( d ) and low expression of CD45 ( e ), as features of myeloid precursor cells. <t>p-STAT5</t> response was then measured on these selected cells by dual SSC/STAT5 cytogram ( f ). In panel ( e ), only CD33+/CD34+/CD14− gated cells are shown. In panel ( f ), only CD33+/CD34+/CD14−/CD45low gated cells are shown.
Anti Phospho Stat5 (P Stat5) Alexa 488, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-phospho-stat5 (p-stat5) alexa 488/product/Becton Dickinson
Average 90 stars, based on 1 article reviews
anti-phospho-stat5 (p-stat5) alexa 488 - by Bioz Stars, 2026-03
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anti-pstat5 monoclonal antibody mab c11c5  (Biocare Medical)
90
Biocare Medical anti-pstat5 monoclonal antibody mab c11c5
Specification of antibodies used in this study
Anti Pstat5 Monoclonal Antibody Mab C11c5, supplied by Biocare Medical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-pstat5 monoclonal antibody mab c11c5/product/Biocare Medical
Average 90 stars, based on 1 article reviews
anti-pstat5 monoclonal antibody mab c11c5 - by Bioz Stars, 2026-03
90/100 stars
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pstat5 (tyr694) antibody  (GeneTex)
90
GeneTex pstat5 (tyr694) antibody
Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and <t>pSTAT5,</t> two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-X L and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. β-Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. β-Actin confirms equal loading of the proteins.
Pstat5 (Tyr694) Antibody, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pstat5 (tyr694) antibody/product/GeneTex
Average 90 stars, based on 1 article reviews
pstat5 (tyr694) antibody - by Bioz Stars, 2026-03
90/100 stars
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alexa-fluor-647–conjugated pstat-5 ab  (Becton Dickinson)
90
Becton Dickinson alexa-fluor-647–conjugated pstat-5 ab
Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and <t>pSTAT5,</t> two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-X L and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. β-Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. β-Actin confirms equal loading of the proteins.
Alexa Fluor 647–Conjugated Pstat 5 Ab, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/alexa-fluor-647–conjugated pstat-5 ab/product/Becton Dickinson
Average 90 stars, based on 1 article reviews
alexa-fluor-647–conjugated pstat-5 ab - by Bioz Stars, 2026-03
90/100 stars
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pstat5 antibody  (Merck & Co)
90
Merck & Co pstat5 antibody
Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and <t>pSTAT5,</t> two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-X L and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. β-Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. β-Actin confirms equal loading of the proteins.
Pstat5 Antibody, supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pstat5 antibody/product/Merck & Co
Average 90 stars, based on 1 article reviews
pstat5 antibody - by Bioz Stars, 2026-03
90/100 stars
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pstat5 antibody  (Active Motif)
90
Active Motif pstat5 antibody
Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and <t>pSTAT5,</t> two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-X L and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. β-Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. β-Actin confirms equal loading of the proteins.
Pstat5 Antibody, supplied by Active Motif, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pstat5 antibody/product/Active Motif
Average 90 stars, based on 1 article reviews
pstat5 antibody - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier
alexa 647–conjugated pstat-5 antibodies  (Becton Dickinson)
90
Becton Dickinson alexa 647–conjugated pstat-5 antibodies
Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and <t>pSTAT5,</t> two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-X L and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. β-Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. β-Actin confirms equal loading of the proteins.
Alexa 647–Conjugated Pstat 5 Antibodies, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/alexa 647–conjugated pstat-5 antibodies/product/Becton Dickinson
Average 90 stars, based on 1 article reviews
alexa 647–conjugated pstat-5 antibodies - by Bioz Stars, 2026-03
90/100 stars
  Buy from Supplier

Image Search Results


Flow cytometric gating strategy adopted to identify CD33+/CD34+ precursor cells. A representative JMML patient is showed. Mononuclear cells were initially gated to exclude debris and residual granulocytes by physical parameters ( a ); all myeloid cells were selected by their reactivity to anti-CD33 antibody ( b ); myeloid precursors were then identified as CD33+/CD34+ double positive cells ( c ). CD33+/CD34+ cells were further checked for their negativity to anti-CD14 antibody ( d ) and low expression of CD45 ( e ), as features of myeloid precursor cells. p-STAT5 response was then measured on these selected cells by dual SSC/STAT5 cytogram ( f ). In panel ( e ), only CD33+/CD34+/CD14− gated cells are shown. In panel ( f ), only CD33+/CD34+/CD14−/CD45low gated cells are shown.

Journal: Blood Cancer Journal

Article Title: Validation of flow cytometric phospho-STAT5 as a diagnostic tool for juvenile myelomonocytic leukemia

doi: 10.1038/bcj.2013.56

Figure Lengend Snippet: Flow cytometric gating strategy adopted to identify CD33+/CD34+ precursor cells. A representative JMML patient is showed. Mononuclear cells were initially gated to exclude debris and residual granulocytes by physical parameters ( a ); all myeloid cells were selected by their reactivity to anti-CD33 antibody ( b ); myeloid precursors were then identified as CD33+/CD34+ double positive cells ( c ). CD33+/CD34+ cells were further checked for their negativity to anti-CD14 antibody ( d ) and low expression of CD45 ( e ), as features of myeloid precursor cells. p-STAT5 response was then measured on these selected cells by dual SSC/STAT5 cytogram ( f ). In panel ( e ), only CD33+/CD34+/CD14− gated cells are shown. In panel ( f ), only CD33+/CD34+/CD14−/CD45low gated cells are shown.

Article Snippet: Samples were incubated with anti-phospho-STAT5 (p-STAT5) Alexa 488 (Y694, clone 47, BD Biosciences) or isotype IgG 1 k Alexa 488 (clone MOPC-21, BD Biosciences), anti-CD33 PE (clone P67.6, BD Biosciences), anti-CD34 APC (clone 8G12, BD Biosciences), anti-CD45 PerCP (clone 2D1, BD Biosciences) and anti-CD38 PE-Cy7 (clone HB7, BD Biosciences) antibodies.

Techniques: Expressing

Training set samples assessed the best threshold. The p-STAT5 responses were measured at each GM-CSF concentration in the training set of samples (11 JMMLs and 23 controls). The best dose to distinguish JMML from non-JMML samples was identified at 0.1 ng/ml of GM-CSF ( P <0.0001). p-STAT5-positive cells (%) were quantified by scaling the maximum % of p-STAT5+ cells at 100 and the unstimulated p-STAT5+ cells to 0.

Journal: Blood Cancer Journal

Article Title: Validation of flow cytometric phospho-STAT5 as a diagnostic tool for juvenile myelomonocytic leukemia

doi: 10.1038/bcj.2013.56

Figure Lengend Snippet: Training set samples assessed the best threshold. The p-STAT5 responses were measured at each GM-CSF concentration in the training set of samples (11 JMMLs and 23 controls). The best dose to distinguish JMML from non-JMML samples was identified at 0.1 ng/ml of GM-CSF ( P <0.0001). p-STAT5-positive cells (%) were quantified by scaling the maximum % of p-STAT5+ cells at 100 and the unstimulated p-STAT5+ cells to 0.

Article Snippet: Samples were incubated with anti-phospho-STAT5 (p-STAT5) Alexa 488 (Y694, clone 47, BD Biosciences) or isotype IgG 1 k Alexa 488 (clone MOPC-21, BD Biosciences), anti-CD33 PE (clone P67.6, BD Biosciences), anti-CD34 APC (clone 8G12, BD Biosciences), anti-CD45 PerCP (clone 2D1, BD Biosciences) and anti-CD38 PE-Cy7 (clone HB7, BD Biosciences) antibodies.

Techniques: Concentration Assay

Representative flow cytometric contour plots of p-STAT5 response in CD33+/CD34+ cells. Dual SSC/STAT5 cytograms from a JMML (upper panels) and a control (lower panels) are shown. Contour plots are referred to CD33+/CD34+ cells identified by gating strategy described in . For each dose of GM-CSF, the raw percentage of responding p-STAT5-positive cells is shown. Response to stimulation at each GM-CSF dose was then quantified by scaling the maximum percentage of p-STAT5+ cells at 100 and the unstimulated p-STAT5+ cells to 0. According to this criteria, calculated p-STAT5 responses are indicated in parenthesis for each stimulation dose.

Journal: Blood Cancer Journal

Article Title: Validation of flow cytometric phospho-STAT5 as a diagnostic tool for juvenile myelomonocytic leukemia

doi: 10.1038/bcj.2013.56

Figure Lengend Snippet: Representative flow cytometric contour plots of p-STAT5 response in CD33+/CD34+ cells. Dual SSC/STAT5 cytograms from a JMML (upper panels) and a control (lower panels) are shown. Contour plots are referred to CD33+/CD34+ cells identified by gating strategy described in . For each dose of GM-CSF, the raw percentage of responding p-STAT5-positive cells is shown. Response to stimulation at each GM-CSF dose was then quantified by scaling the maximum percentage of p-STAT5+ cells at 100 and the unstimulated p-STAT5+ cells to 0. According to this criteria, calculated p-STAT5 responses are indicated in parenthesis for each stimulation dose.

Article Snippet: Samples were incubated with anti-phospho-STAT5 (p-STAT5) Alexa 488 (Y694, clone 47, BD Biosciences) or isotype IgG 1 k Alexa 488 (clone MOPC-21, BD Biosciences), anti-CD33 PE (clone P67.6, BD Biosciences), anti-CD34 APC (clone 8G12, BD Biosciences), anti-CD45 PerCP (clone 2D1, BD Biosciences) and anti-CD38 PE-Cy7 (clone HB7, BD Biosciences) antibodies.

Techniques:

Comparison of p-STAT5-positive cells (%) induced by 0.1 ng/ml of GM-CSF in the validation series comprising 11 JMML samples (central box plot), 24 controls (left box plot) and 7 samples from patients with other diseases mimicking JMML at presentation (right box plot). The discriminating threshold (17.17% as assessed in the training set) is indicated. The bold line inside each box plot indicates the median level, while the upper and lower lines indicate the maximum and minimum observed values, respectively. There are no outliers.

Journal: Blood Cancer Journal

Article Title: Validation of flow cytometric phospho-STAT5 as a diagnostic tool for juvenile myelomonocytic leukemia

doi: 10.1038/bcj.2013.56

Figure Lengend Snippet: Comparison of p-STAT5-positive cells (%) induced by 0.1 ng/ml of GM-CSF in the validation series comprising 11 JMML samples (central box plot), 24 controls (left box plot) and 7 samples from patients with other diseases mimicking JMML at presentation (right box plot). The discriminating threshold (17.17% as assessed in the training set) is indicated. The bold line inside each box plot indicates the median level, while the upper and lower lines indicate the maximum and minimum observed values, respectively. There are no outliers.

Article Snippet: Samples were incubated with anti-phospho-STAT5 (p-STAT5) Alexa 488 (Y694, clone 47, BD Biosciences) or isotype IgG 1 k Alexa 488 (clone MOPC-21, BD Biosciences), anti-CD33 PE (clone P67.6, BD Biosciences), anti-CD34 APC (clone 8G12, BD Biosciences), anti-CD45 PerCP (clone 2D1, BD Biosciences) and anti-CD38 PE-Cy7 (clone HB7, BD Biosciences) antibodies.

Techniques:

Comparison of p-STAT5-positive cells (%) induced by 0.1 ng/ml of GM-CSF according to the different cell source. In all, 17 JMML BM samples (left box plot), 5 JMML PB samples (middle left box plot), 12 BM samples and 2 PB samples (middle right and right box plot, respectively) from patients with other diseases mimicking JMML are shown. The discriminating threshold (17.17% as assessed in the training set) is indicated. The bold line inside each box plot indicates the median level, while the upper and lower lines indicate the maximum and minimum observed values, respectively. There are no outliers.

Journal: Blood Cancer Journal

Article Title: Validation of flow cytometric phospho-STAT5 as a diagnostic tool for juvenile myelomonocytic leukemia

doi: 10.1038/bcj.2013.56

Figure Lengend Snippet: Comparison of p-STAT5-positive cells (%) induced by 0.1 ng/ml of GM-CSF according to the different cell source. In all, 17 JMML BM samples (left box plot), 5 JMML PB samples (middle left box plot), 12 BM samples and 2 PB samples (middle right and right box plot, respectively) from patients with other diseases mimicking JMML are shown. The discriminating threshold (17.17% as assessed in the training set) is indicated. The bold line inside each box plot indicates the median level, while the upper and lower lines indicate the maximum and minimum observed values, respectively. There are no outliers.

Article Snippet: Samples were incubated with anti-phospho-STAT5 (p-STAT5) Alexa 488 (Y694, clone 47, BD Biosciences) or isotype IgG 1 k Alexa 488 (clone MOPC-21, BD Biosciences), anti-CD33 PE (clone P67.6, BD Biosciences), anti-CD34 APC (clone 8G12, BD Biosciences), anti-CD45 PerCP (clone 2D1, BD Biosciences) and anti-CD38 PE-Cy7 (clone HB7, BD Biosciences) antibodies.

Techniques:

Specification of antibodies used in this study

Journal: Veterinary and comparative oncology

Article Title: The JAK2/STAT5 signaling pathway as a potential therapeutic target in canine mastocytoma

doi: 10.1111/vco.12311

Figure Lengend Snippet: Specification of antibodies used in this study

Article Snippet: Immunohistochemistry (IHC) was performed on sections prepared from paraffin-embedded, formalin-fixed mastocytoma specimens by the indirect immune-peroxidase staining technique using the anti-pSTAT5 monoclonal antibody (mAb) C11C5 (diluted 1:20 in Renoir Red Diluent, Biocare Medical, Walnut Creek, California) as described.

Techniques: Transduction

Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and pSTAT5, two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-X L and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. β-Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. β-Actin confirms equal loading of the proteins.

Journal: Journal of Cellular and Molecular Medicine

Article Title: Inhibition of IGF-IR tyrosine kinase induces apoptosis and cell cycle arrest in imatinib-resistant chronic myeloid leukaemia cells

doi: 10.1111/j.1582-4934.2009.00795.x

Figure Lengend Snippet: Effects of inhibition of IGF-IR on IGF-IR, BCR-ABL and downstream target proteins in CML cell lines. (A) PPP induces concentration-dependent decrease in IGF-IR tyrosine kinase activity in K562 and KBM-5 cell lines. In contrast, PPP fails to cause similar effect in BCR-ABL tyrosine kinase activity. The results represent the means ± S.D. of three consistent experiments. *: P < 0.01 and †: P < 0.001 compared with control untreated cells. (B) Western blotting and co-immunoprecipitation studies confirm that PPP decreases the tyrosine phosphorylation of IGF-IR in a concentration-dependent fashion (results shown are representative and were obtained from the KBM-5 cell line). The basal levels of IGF-IR did not change after treatment with PPP. The phosphorylation level of BCR-ABL remains unchanged after treatment with PPP. The decrease in pIGF-IR is associated with down-regulation of pAkt and pSTAT5, two oncogenic proteins in CML. Changes are not seen in Akt and STAT5. Also, PPP induces changes consistent with apoptotic cell death including down-regulation of Bcl-2, Bcl-X L and caspase-3. Moreover, treatment with PPP induces up-regulation of cyclin B1 and down-regulation of cyclin E and pCdc2, whereas the levels of Cdc2 and p16 remain unchanged. Overall, the changes in the cell cycle regulatory proteins are consistent with G2/M-phase cell cycle arrest. β-Actin shows equal loading of the proteins. (C) IGF-IR siRNA decreases IGF-IR levels in the KBM-5 cell line and this decrease is associated with down-regulation of pAkt and pSTAT5. Basal levels of these two proteins remain unchanged. IGF-IR siRNA did not induce alterations in BCR-ABL or pBCR-ABL protein. β-Actin confirms equal loading of the proteins.

Article Snippet: Antibodies obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) included Bcl-2 (catalogue number: sc-7382), cyclin B1 (sc-7393), cyclin E (sc-198), Cdc2 (sc-52316), pCdc2 (Thr14/Tyr15; sc-12340-R) and p16 (sc-56330); from Cell Signaling Technology (Danvers, MA, USA) were pIGF-IR (Tyr1131; 3021), pBCR-ABL (p-c-Abl; Tyr412; 2865), Akt (9272) and pAkt (Ser473; 587F11); from Zymed Laboratories (South San Francisco, CA, USA) were IGF-IRβ (39–6700) and Bcl-X L (18–0217); from Calbiochem (Gibbstown, NJ, USA) was BCR-ABL (c-Abl; OP19); from R&D Systems (Minneapolis, MN, USA) was STAT5 (MAB2174); from GeneTex Incorporation (San Antonio, TX, USA) was pSTAT5 (Tyr694; GTX52364) and from Sigma (St. Louis, MO, USA) was β-Actin (A-2228).

Techniques: Inhibition, Concentration Assay, Activity Assay, Western Blot, Immunoprecipitation