Figure 1.

DHX16 interacts with unanchored poly-Ub chains

(A) Unanchored Ub enrichment from murine lung tissue.

(B) DEAH-box RNA helicases identified from murine lungs as unanchored Ub-interacting factors (MS).

(C) Unanchored Ub enrichment utilizing GST-UBD-IsoT.

(D) Interaction between GST-UBD-IsoT and recombinant K48-poly-Ub chains (2-16 Ub/chain) in an in vitro Ub-binding assay (coIP).

(E) GST-UBD-IsoT was mixed with lung lysates from mice treated with NTC or TRIM6 PPMO followed by GST pulldown (lung samples used in Rajsbaum et al., 2014b) (coIP).

(F) Interaction between endogenous DHX16 and GST-UBD-IsoT from HEK293Ts (coIP).

(G) DHX16 interacts non-covalently with poly-Ub after normal (150 mM NaCl), but not high salt (550 mM NaCl) washes (denaturing coIP).

(H) DHX16 interacts non-covalently with poly-Ub after RIPA, but not urea washes. Arrows indicate DHX15 interactions with either non-covalent or covalent poly-Ub (denaturing coIP).

(I) DHX16 interacts with K48-poly-Ub in an in vitro Ub binding assay (coIP).

(J) Endogenous DHX16 interacts non-covalently with K48-poly-Ub after normal, but not high salt washes (denaturing coIP).

(K) DHX16 interacts with K48-poly-Ub in HEK293Ts infected with IAV for 12 hrs (PR8 MOI = 5) (coIP).

Data are representative of 2–3 independent experiments.

See also Figure S1.