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. 2015 Jul 6;112(32):10014–10019. doi: 10.1073/pnas.1507534112

Fig. 1.

Fig. 1.

Ube2W and Ube2N are required for TRIM21-mediated signaling and neutralization. (A) Proteasome or NF-κB signaling inhibitors prevent NF-κB activation in response to AdV/IgG. (B and C) TE671 cells, infected with AdV (MOI ∼0.1), preincubated with 9C12 or PBS, treated with the inhibitors indicated or DMSO (B), or expressing scrambled shRNA (shScr) (C–G), Ube2N-specific shRNA (shUbe2N) (C and F) or Ube2W-specific shRNA (shUbe2W) (C–E and G), percentage infection quantified by flow cytometry for GFP expression 24 h postinfection, values normalized to PBS-treated AdV control. (D and E) TE671-NF-κB-Luc cells expressing shScr or shUbe2W challenged with PBS (D and E), human IgG, AdV, AdV/IgG (D), or human TNFα (E) for 7 h. (F) Immunoblot of TE671-shScr or TE671-shUbe2N, detecting Ube2N or β-actin (loading control). (G) Quantitative RT-PCR detecting Ube2W mRNA in TE671-shScr or TE671-shUbe2W. Values calculated using the ∆∆Ct method and normalized to β-Actin mRNA. All error bars are SEM of duplicates or triplicates.