Nucleo-Cytoplasmic Trafficking of TRIM8, a Novel Oncogene, Is Involved in Positive Regulation of TNF Induced NF-κB Pathway
Figure 2
Cellular localization of different domains of TRIM8 and their role in NF-κB activation.
(A) Schematic diagram showing TRIM8 domains and constructs used for transfection. (B) Cellular localization of TRIM8 deletion constructs. HeLa cells were plated on cover slip and transfected with GFP-tagged different deletion constructs of TRIM8. After 24 hours of transfection, the cells were fixed with 4% paraformaldehyde, stained with DAPI and visualized by confocal microscopy. Scale bar represent 7.5 µM. (C) RING domain is essential for positive regulation of TNF induced NF-κB activation. HEK293 was co-transfected with full length TRIM8, ΔRING, ΔB1, ΔCC, ΔC-ter, vector, treated with TNFα and NF-κB activation measured by Dual Glo luciferase assay. (D) RING, CC domain and C-terminal region of TRIM8 is essential for p65 mediated NF-κB activity. HEK293 was co-transfected with full length TRIM8, ΔRING, ΔB1, ΔCC, ΔC-ter, vector with p65-GFP and NF-κB activation measured by Dual Glo luciferase assay. Asterisk (*) indicates NF-κB activation significantly changed between groups: p value<0.05 for SEM of minimum three independent experiments.