FIG 5.

Both SNW1 and p65 complex with p-TEFb upon TNF-α treatment. THP-1 cells were treated with TNF-α for 30 or 60 min and immunoprecipitated for either SNW1 (A) or p65 (B). (A) Constitutive binding of p-TEFb (CDK9) to SNW1 inside THP-1 cells, with significantly increased binding to RNA Pol II, along with p65, upon TNF-α treatment. (B) Increased binding of p65 to p-TEFb and SNW1 upon treatment with TNF-α, as expected. The binding of IκBα and p50 to p65 is used as a positive control, while the lack of binding of actin (a ubiquitous highly expressed protein) to either SNW1 or p65 is used as a negative control. Note that for panel A, SNW1, Pol II, and p-TEFb were blotted on the same gel, while p65 and actin were blotted using the same samples but on a different gel (due to similar molecular weights of p65, SNW1, p-TEFb, and actin). Similarly, for panel B, p65, p-TEFb, and IκBα were blotted on the same gel, while SNW1, p50, and actin were blotted using the same samples but on a different gel.