anti psyk  (Cell Signaling Technology Inc)

Journal: Redox Biology

Article Title: ROS via BTK-p300-STAT1-PPARγ signaling activation mediates cholesterol crystals-induced CD36 expression and foam cell formation

doi: 10.1016/j.redox.2016.12.005

Figure Lengend Snippet: CC induces p300 activation in BTK-dependent manner. A. Equal amounts of protein from control and the indicated time periods of CC (40 μg/ml)-treated cells were immunoprecipitated with anti-p300 antibodies or IgG and the immunocomplexes were analyzed by Western blotting with anti-PY20 or anti-phosphoserine/threonine antibodies followed by normalization for p300. B. All the conditions were same as in panel A except that the cell extracts were analyzed by Western blotting for pBTK, pPyk2, pSrc and pSyk levels using their phospho-specific antibodies and normalized for their total levels. C. Cells were transfected with the indicated ASO, quiesced, treated with and without CC and analyzed for CD36 expression, oxLDL uptake or foam cell formation as described in , panel C. The blot was reprobed for BTK, Pyk2 and β-tubulin levels to show the effects of the ASOs on their target and off target molecules levels. D. Cells were transfected with control or BTK ASO, quiesced, treated with and without CC for 1 h and cell extracts were prepared. Equal amount of protein from control and each treatment were immunoprecipitated with anti-p300 or anti-STAT1 antibodies and the immunocomplexes were analyzed by Western blotting for anti-PY20, anti-STAT1, anti-Ac-Lys or anti-PPARγ antibodies and normalized for p300 or STAT1. An equal amount of protein from control and each treatment was also assayed for p300 acetyltransferase activity. *p<0.01 vs control ASO; **p<0.01 vs control ASO+CC.

Article Snippet: Anti-pBTK (5082), anti-pPyk2 (3291), anti-pSrc (2101), anti-pSTAT1 (9177), anti-pSTAT2 (4441), anti-pSTAT3 (9133), anti-pSTAT4 (5267), anti-pSTAT5 (9351), anti-pSTAT6 (9364), and anti-pSyk (2715) antibodies were bought from Cell Signaling Technology (Beverly, MA).

Techniques: Activation Assay, Control, Immunoprecipitation, Western Blot, Transfection, Expressing, Activity Assay