Figure 1.

JAK1 interacts Sirt1. A, JAK1 expression plasmid was co-transfected with or without FLAG-Sirt1 into HEK293 cells. The interaction of Sirt1 with JAK1 in the transfected cells was analyzed by co-immunoprecipitation with anti-FLAG antibody and by Western blotting with JAK1 antibody (top panel). The same membrane was reprobed with anti-Sirt1 (middle panel). JAK1 expression in the whole cell lysate was determined by Western blotting (bottom panel). B–D, MCF-10 (B), HCT116 (C), and MM.1s (D) cells were stimulated with or without 10 ng/ml IL-6 and lysed with RIPA buffer. The interaction of Sirt1 and JAK1 was determined by immunoprecipitation of JAK1 using normal rabbit IgG (rIgG) as a control and by Western blotting with anti-Sirt1 antibody (top panels). The same membrane was stripped and reblotted with anti-JAK1 (middle panels). Sirt1 expression in the whole cell lysate was determined by Western blotting (bottom panels).