Fig. 2
From: Acetylation of AGO2 promotes cancer progression by increasing oncogenic miR-19b biogenesis
P300/CBP acetylate AGO2. a Serum stimulates AGO2 acetylation. 293T cells were serum-starved for 24 h and followed by stimulation with 20% serum for 1, 2, 3 h; AGO2 acetylation was measured by IP/WB. b P300 and CBP interact with AGO2. HA-P300 or HA-CBP was co-transfected with or without Myc-AGO2 into 293T cells. The association between AGO2 and P300 or CBP was determined by IP/WB. Also see Figure S2B. c Overexpression of P300/CBP significantly increases AGO2 acetylation. Myc-AGO2 with HA-P300 or HA-CBP were transfected into 293T cells. AGO2 acetylation was determined by IP with anti-Myc antibody (left panel) or anti-Ac antibody (right panel), and followed by WB. d Knockdown of P300 attenuates AGO2 acetylation. P300-siRNA (40 nM) was transfected into 293T cells for 48 h, and AGO2 acetylation were examined by IP/WB. e P300 significantly increases acetylation of AGO2-WT rather than AGO2-3KR. Myc-tagged AGO2-WT or AGO2-3KR was co-transfected with or without HA-P300 into 293T cells, and then AGO2 acetylation was analyzed by IP with anti-Myc antibody and WB with anti-Ac antibody. f P300 mediates AGO2 acetylation at K355, K493 and K720. Flag-tagged AGO2 was co-transfected with or without HA-P300 into 293T cells, and AGO2 acetylation was analyzed by IP with anti-Flag antibody and WB with AGO2 specific acetyl-antibodies. g, h P300 increases AGO2 acetylation in vitro. Purified GST-AGO2 protein was incubated in cell lysates of 293T cells expressing HA-P300 or the control vector. Acetylation of GST-AGO2 was analyzed by GST pull down and followed by WB with anti-Ac antibody (g) or AGO2 specific acetyl-antibodies (h)
