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Figure 4: K7 reduces vGPCR protein expression.(A) K7 reduces vGPCR protein expression in a dose-dependent manner. Human endothelial ECV cells were transfected with plasmids containing vGPCR-Flag and K7-V5 as indicated. WCLs were analyzed by immunoblot with anti-Flag (vGPCR, top panel), anti-V5 (K7, middle panel), and anti-tubulin (bottom panel) antibodies. (B) The putative K7 TM domain is necessary for vGPCR downregulation. Transfection of ECV cells and immunoblot analyses were performed as in (A). Data represents three independent experiments. (C) K7 reduces vGPCR-mediated activation of NF-κB, NF-AT, and AP-1 transcription factors. 293T cells were transfected with reporter plasmid cocktail, and plasmids containing vGPCR and K7. Luciferase activity normalized against β-galactosidase activity is shown. Error bars denote standard deviation and data represent three independent experiments. (D) K7 knockdown by shRNA-mediated silencing. (Left Top) The relative genomic locations of K7 coding sequence and the transcribed region of PAN were shown. The numbers indicate the nucleotide position according to a published KSHV genome sequence (accession number: U75698). Bars represent relative location of sequences base paired with four shRNAs within the 5′ untranslated region of K7. The order of shRNAs on the diagram is: shRNA#2, #3, #1, and #4. (Left Bottom) A diagram shows the experimental design of lentivirus infection and KSHV lytic replication induced by TPA. (Right) RT-PCR analyses were performed using gene specific primers for K7, β-actin, vGPCR, and PAN as described in Materials and Methods. Numbers indicate intensity of K7 band measured by densitometry. Data represent two independent experiments.
Image Text (High Precision): 100 2002 AP-1 AT Activation K7-V5 NF PAN infection lentivirus
Other Images from "Kaposi's Sarcoma-Associated Herpesvirus K7 Induces Viral G Protein-Coupled Receptor Degradation and Reduces Its Tumorigenicity":
Figure 6 K7 induces proteasome-dependent degradat...
Figure 5 Increased vGPCR protein expression by sh...
Figure 3 Overlapped expression of K7 and vGPCR du...
Figure 2 Intracellular localization of vGPCR and ...
Figure 1 KSHV K7 interacts with vGPCR.(A) K7 inte...
Figure 7 K7 retains vGPCR in the ER.(A) vGPCR loc...
Figure 8 The effect of vGPCR and K7 on cell growt...
Figure 9 K7 negatively regulates vGPCR tumorigeni...
Figure 4 K7 reduces vGPCR protein expression.(A) ...