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. Author manuscript; available in PMC: 2024 Feb 5.
Published in final edited form as: Cell Rep. 2023 Oct 8;42(10):113223. doi: 10.1016/j.celrep.2023.113223

Figure 6. SUGP1 recruits DHX15 via its ULM domain.

Figure 6.

(A) Diagram of SUGP1’s primary domain structure.

(B) Protein blots and quantification of biotinylated proteins labeled by FL versus truncated SUGP1-DHX15 split-APEX activity. Ponceau S protein stain, loading control for total protein; streptavidin IRDye, detection of biotinylated proteins; FRB-nls, FRB control protein fused with an SV40 NLS; ΔGp, G-patch domain truncation; ΔULM, ULM truncation; ΔS1&2, SURP1 and SURP2 truncation; colored asterisks, corresponding bands to FL and truncated HA-SUGP1-EX.

(C) Protein blots of biotinylated proteins labeled in (B) enriched by streptavidin pull-down experiments.

(D) Quantification of pull-downs in (C).

(E) Fluorescent microscopy images of biotinylation signals (streptavidin, magenta), antibodies detecting HA-tagged SUGP1-EX, FL versus truncations (HA, red), FLAG-tagged AP-DHX15 (FLAG, green), nuclear DNA dye (DAPI, blue), and merged channels.

(F) Fluorescent microscopy images of antibody detecting HA-tagged SUGP1.W387A-EX (HA, red), nuclear DNA dye (DAPI, blue), and merged channels.

(G) Protein blots and quantification of biotinylated proteins labeled by full-length (FL), ΔULM, versus W387A mutant SUGP1-DHX15 interaction-reconstituted split-APEX activity. Ponceau S protein stain, loading control for total protein; streptavidin IRDye, detection of biotinylated proteins. Biotinylated HA was detected by merging the streptavidin channel (green) with the HA channel (red). See also Figure S4.