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. 2016 Jul 29;36(16):2182–2194. doi: 10.1128/MCB.00174-16

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FIG 1 — Fbl12 promotes p21 ubiquitination. (A) Coimmunoprecipitation of Myc-Fbl12 and Flag-p21 in HEK 293T cells. (B) GST pulldown of His-Fbl12/Skp1 and either GST or GST-p21. The precipitated proteins were analyzed by immunoblotting. (C, top) Schematic structure of Fbl12. F-Box, F-box domain; LRR, leucine-rich repeat. (Bottom) Coimmunoprecipitation of p21 and either EGFP-Fbl12, EGFP-Fbl12 F-box, or EGFP-Fbl12ΔF. (D, top) Schematic structure of p21. CDI, CDK inhibitor domain; NLS, nuclear localization signal. (Bottom) Coimmunoprecipitation of Flag-Fbl12 and either EGFP-p21, EGFP-p21ΔNLS, or EGFP-p21NLS. (E) Ubiquitinated proteins were purified from denatured cell lysates using Talon metal affinity resin and analyzed by immunoblotting.