src kinase inhibitor 1 ski 1  (Santa Cruz Biotechnology)

Journal: Journal of Inflammation (London, England)

Article Title: Different roles for non-receptor tyrosine kinases in arachidonate release induced by zymosan and Staphylococcus aureus in macrophages

doi: 10.1186/1476-9255-3-8

Figure Lengend Snippet: Inhibition by PP2 or SKI-1 of zymosan- and S.aureus -induced release of arachidonate . Macrophages were labeled with [3H]arachidonic acid for 20 h. The cells were pretreated for 15 min with the indicated concentrations of PP2 (A) or SKI-1 (B) followed by stimulation with either zymosan (●) for 45 min or S.aureus (▲) for 60 min. Results are mean ± SEM (n = 3) and corrected for the release in control cultures.

Article Snippet: LFM-A13 and Src kinase inhibitor I (SKI-1) from Calbiochem (La Jolla, CA, USA).

Techniques: Inhibition, Labeling

Journal: Journal of Inflammation (London, England)

Article Title: Different roles for non-receptor tyrosine kinases in arachidonate release induced by zymosan and Staphylococcus aureus in macrophages

doi: 10.1186/1476-9255-3-8

Figure Lengend Snippet: Inhibition by PP2 or SKI-1 of zymosan- and S.aureus - induced phosphorylation of MAP kinases . Macrophages were pretreated for 15 min with PP2 (1–10 μM), followed by stimulation with zymosan ( A ) or S.aureus ( B ) for 20 min. ( C ) Macrophages were pretreated for 15 min with SKI-1 (5 μM), followed by stimulation with zymosan or S.aureus for 20 min. Equal amounts of cell lysate were run on 10% polyacrylamide gels and probed with phosphospecific antibodies against ERK, p38 and JNK. The membrane was reprobed with ERK-2 antibody to verify equal loading of protein.

Article Snippet: LFM-A13 and Src kinase inhibitor I (SKI-1) from Calbiochem (La Jolla, CA, USA).

Techniques: Inhibition

Journal: Journal of Inflammation (London, England)

Article Title: Different roles for non-receptor tyrosine kinases in arachidonate release induced by zymosan and Staphylococcus aureus in macrophages

doi: 10.1186/1476-9255-3-8

Figure Lengend Snippet: Zymosan but not S.aureus induced tyrosine phosphorylation of PLC γ2 . ( A ) Macrophages were stimulated with zymosan(z) or S.aureus ( S.a ) for 45 min. (B and C) Macrophages were pretreated for 15 min with either PP2 (5 μM), wortmannin (W, 100 nM) ( B ) or SKI-1 (5 μM) ( C ) followed by stimulation with zymosan for 30 min. Cell lysates were immunoprecipitated with antibody against PLCγ2 as described, followed by Western blot analysis with phosphotyrosine-specific antibody. The membrane was stripped and reprobed with antibody against PLCγ2. The data are representative of three separate experiments.

Article Snippet: LFM-A13 and Src kinase inhibitor I (SKI-1) from Calbiochem (La Jolla, CA, USA).

Techniques: Immunoprecipitation, Western Blot

Journal: Journal of Inflammation (London, England)

Article Title: Different roles for non-receptor tyrosine kinases in arachidonate release induced by zymosan and Staphylococcus aureus in macrophages

doi: 10.1186/1476-9255-3-8

Figure Lengend Snippet: Zymosan and bacteria but not LPS or peptidoglycan (PGN) induce phosphorylation of Btk . ( A ) Macrophages were stimulated with zymosan (zym), S.aureus ( S.a .), LPS or PGN for 30 min. (B and C) Macrophages were pretreated for 15 min with either wortmannin (W, 100 nM), PP2 (5 μM), SU6656 (5 μM) or SKI-1 (5 μM), followed by stimulation with zymosan ( B ) or S.aureus ( C ) for 30 min. Equal amounts of cell lysate were run on polyacrylamide gels and probed with phosphospecific antibodies against Btk. The membrane was reprobed with ERK-2 antibody to verify equal loading of proteins. The data are representative of three separate experiments.

Article Snippet: LFM-A13 and Src kinase inhibitor I (SKI-1) from Calbiochem (La Jolla, CA, USA).

Techniques:

Journal: PLoS ONE

Article Title: The Ubiquitin Ligase CBLC Maintains the Network Organization of the Golgi Apparatus

doi: 10.1371/journal.pone.0138789

Figure Lengend Snippet: Giantin staining in HeLa cells. Fragmentation Index measured in quadruplicates on at least 400 cells per condition. Error bars show SD statistical significance (p) measured by unpaired Student’s t-test. (*) represents p<0.05, (**) represents p<0.01 and (***) represents p<0.001. (A and D) Co-transfection of CBLC and SRC siRNAs in HeLa cells (B and E) Treatment of CBLC-depleted cells with SRC inhibitors PP2 or SKI-1 rescues Golgi morphology. (C and F) Co-transfection of CBLC and ARF1 siRNAs in HeLa cells rescues Golgi morphology. Scale bar = 10 μm.

Article Snippet: After 48 hours of siRNA transfection, medium was aspirated, replaced with fresh medium containing 5 μM SRC inhibitors PP2 or SRC inhibitor kinase 1 SKI-1 (Sigma) and incubated at 37°C in a 10% CO 2 humidified incubator for a further 24 hours before fixation.

Techniques: Staining, Cotransfection