Extended Data Fig. 5: WEE1 is acetylated at Lys177.
From: SIRT1 deacetylates WEE1 and sensitizes cancer cells to WEE1 inhibition
(a) The sequence of the region containing the acetylation site of WEE1 was aligned with those from various species. (b) Characterization of the anti-AcK177-WEE1 antibody by a dot blot assay. Various amounts of unacetylated or acetylated WEE1-K177 peptides were spotted onto nitrocellulose membrane and immunoblotted with the anti-AcK177-WEE1 antibody. (c) HEK293T cells were transfected with the indicated plasmids for 24 hr. Cell lysates were then prepared and western blot analysis was carried out as indicated. (d) HEK293T cells were transfected with the indicated plasmids for 24 hr. Cells were then left untreated or irradiated with 40 J/m2 UV and allowed to recover for 4 hr. Cell lysates were then prepared and western blot analysis was carried out as indicated. (e) HEK293T cells transfected with the indicated plasmids were left untreated or irradiated with 40 J/m2 UV and allowed to recover for 4 hr. Cell lysates were then prepared and western blot analysis was carried out as indicated. (f) U2OS cells were left untreated or treated with HU (4 mM), CPT (1 μM), or IR (10 Gy) for 4 hr. Cell lysates were then prepared and western blot analysis was carried out as indicated.
