d08  (TargetMol)

Journal: Frontiers in Microbiology

Article Title: CBX4 facilitates EV71 replication by SUMOylation and stabilizing 3D polymerase

doi: 10.3389/fmicb.2026.1775950

Figure Lengend Snippet: SUMOylation inhibitor 2-D08 suppresses EV71 replication. (A) RD cells were added with 0, 20, 50, and 100 μΜ 2-D08 for 24 h, the cell viability was determined by CCK8 assay. The results were shown as percentage relative to control. (B–E) RD or HT29 cells were treated with EV71 (MOI = 1) and 2-D08 (0, 20, 50, 100 μM), after 16 h, the supernatants and cells were collected. The expression levels of CBX4, 3D, VP1, 3C and GAPDH proteins were detected by Western blotting (B) , the mRNA level of VP1 was determined by qPCR assay (C) , the viral titers in supernatants from RD (D) or HT29 (E) cells were measured by plaque assay. (F) HEK293T cells were transfected with HA-3D plasmid for 6 h, and then treated with different concentrations of 2-D08 (0, 20, 50, 100 μM) for 24 h. The protein levels of HA-3D and GAPDH were detected by immunoblotting. (G) HEK293T cells were transfected with GFP-3D, Myc-SUMO1, HA-Ubc9 and HA-CBX4 plasmids for 6 h, and then treated with 2-D08 (50 μM) for 24 h. The cell lysates were immunoprecipitated with anti-GFP immunomagnetic beads. The IP and input complex were analyzed with indicated antibodies. The graphs are shown as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Inhibitors including UNC3866 (HY-100832), 2-D08 (HY-114166), cycloheximide (CHX) (HY-12320) and MG132 (HY-13259) were purchased from MedchemExpress (Monmouth Junction, NJ, USA).

Techniques: CCK-8 Assay, Control, Expressing, Western Blot, Plaque Assay, Transfection, Plasmid Preparation, Immunoprecipitation

Journal: Frontiers in Microbiology

Article Title: CBX4 facilitates EV71 replication by SUMOylation and stabilizing 3D polymerase

doi: 10.3389/fmicb.2026.1775950

Figure Lengend Snippet: A model illustrating how CBX4 promotes EV71 replication. Host SUMO E3 ligase CBX4 interacts with EV71-encoded 3D polymerase and induces the SUMOylation of 3D, which results in the increased stability of 3D protein and EV71 replication. Pharmacological inhibition of protein SUMOylation with 2-D08 diminishes the SUMOylation and expression of 3D, and ultimately affects the replication of EV71.

Article Snippet: Inhibitors including UNC3866 (HY-100832), 2-D08 (HY-114166), cycloheximide (CHX) (HY-12320) and MG132 (HY-13259) were purchased from MedchemExpress (Monmouth Junction, NJ, USA).

Techniques: Inhibition, Expressing

Journal: bioRxiv

Article Title: Nup358 Sustains Intestinal Epithelial Homeostasis by Preventing Dvl1 Condensate Formation to Restrain Wnt Signaling

doi: 10.64898/2026.03.25.714063

Figure Lengend Snippet: a. Schematic representing different functional domains of Nup358. b. Fluorescent Wnt reporter activity in HEK293T cells transfected with control siRNA, Nup358-specific siRNA, Ubc9-specific siRNA, or treated with the SUMOylation inhibitor 2-D08. c. Schematic representing different HA-tagged truncated forms of Nup358 that were transiently expressed in HEK293T cells. d. Fluorescent Wnt reporter activity in HEK293T cells transfected with either control or Nup358-specific siRNA alone or in combination with different HA-tagged truncated forms of Nup358 were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. ****p ≤ 0.0001 e. Western blot analysis of co-immunoprecipitation of HA-Nup358 and EGFP-Dvl1 transiently expressed in HEK293T cells. GAPDH was used as loading control. f. Immunofluorescence of HEK293T cells transfected with HA-Tagged Nup358 1–1133 and EGFP-Dvl1and stained with an anti-HA antibody. g. Immunofluorescence staining for endogenous Dvl1 and Nup358 in HEK293T cells transfected with either scramble control or Nup358-specific siRNA alone or in combination with HA-Tagged Nup358 1–1133 fragment were analyzed by confocal microscopy (top) and quantified (bottom). Data are expressed as mean ± SD. **p ≤ 0.01.

Article Snippet: For SUMOylation inhibition, cells were treated with 25 μM 2-D08 (Selleckchem, S8696) 72 hours after transfection with either control or Nup358 siRNA.

Techniques: Functional Assay, Activity Assay, Transfection, Control, Confocal Microscopy, Western Blot, Immunoprecipitation, Immunofluorescence, Staining