Journal: Cureus

Article Title: Acute Effects of Growth Hormone on the Cellular Immunologic Landscape in Pediatric Patients

doi: 10.7759/cureus.57383

Figure Lengend Snippet: The 29 antibody panels used to stain PBMCs for CyTOF analysis. PBMCs: Peripheral blood mononuclear cells, CyTOF: Time of flight mass cytometry

Article Snippet: pSTAT5 , 147Sm , Fluidigm , 47 , 3147012A.

Techniques: Staining

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

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