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. 2018 Jul 17;24(3):594–606. doi: 10.1016/j.celrep.2018.06.048

Figure 3.

Figure 3

LUNA Encodes a Cysteine Protease Sensitive to Isopeptidase Inhibitors

(A and B) HFFs transfected with LUNA were incubated with MG132 and analyzed for FLAG (a) and PML expression (b) 24 hr post-transfection (A) and number of ND10- and LUNA-positive cells enumerated (B). Data are mean ± SD and represent triplicate analyses performed in three independent experiments.

(C and D) N-terminal FLAG tagged LUNA (LUNA) transfected into fibroblasts were incubated with isopeptidase inhibitor, G5, and stained for FLAG (a) and PML (b) expression 48 hr later (C) and number of ND10- and LUNA-positive cells enumerated (D). Data are mean ± SD and represent triplicate analyses performed in three independent experiments.

(E) Identification of the two conserved motifs in LUNA (red box). Asterisk denotes catalytic cysteine.

(F and G) N-terminal FLAG tagged catalytic dead mutant of LUNA (LUNAg233c) was transfected into fibroblasts and stained for FLAG (a) and PML (b) expression 48 hr later (F) and the number of ND10- and LUNA-positive cells enumerated (G). Data are mean ± SD and represent triplicate analyses performed in three independent experiments.

See also Figures S2 and S3.