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. 2021 Aug 23;22(16):9094. doi: 10.3390/ijms22169094

Figure 1.

Figure 1

TRIM25 interacts with DDX3X. (A) Anti-FLAG immunoprecipitation (IP) of whole-cell lysates (WCL) of HEK293T cells expressing FLAG-TRIM25 and HA-DDX3X, as well as FLAG-DDX3X and HA-TRIM25. Immunoblot (IB) analysis of IP with anti-FLAG and anti-HA antibodies and WCL with anti-FLAG, anti-HA and anti-actin antibodies. (B) Interaction between endogenous TRIM25 and endogenous DDX3X in HEK293T cells, with immunoblot analysis of WCL and anti-TRIM25 (top) or anti-DDX3X (bottom) IP. (C–E) Anti-FLAG IP of WCL of HEK293T cells overexpressing various recombinant proteins. IB analysis of IP with anti-FLAG and anti-HA antibodies and WCL with anti-FLAG, anti-HA and anti-actin antibodies. (C) FLAG-DDX3X together with HA-tagged wild-type TRIM25, TRIM25ΔRING or TRIM25ΔPRY-SPRY. In the schematic, R = RING, B = B-box, CC = coiled-coil. (D) FLAG-tagged wild-type DDX3X, DDX3X 1–517, DDX3X 1–414 and DDX3X NTE (residues 1–168) constructs together with HA-TRIM25. In the schematic, NTE = N-terminal extension and R = RecA-like domain. (E) FLAG-TRIM25 and HA-tagged wild-type DDX3X or DDX3XΔNTE.