Expression of IRF-7C RNA is associated with type III latency.

<p>(A) Schematic diagram of IRF-7C-specific RPA probe. The bar represents ORF. The IRF-7C-specific RPA probe encompasses the splicing junction. The IRF-7-specific probe region for all splicing variants is also shown. The drawing is not on scale. (B) IRF-7C RNA is highly expressed in cells with EBV type III latency. Total RNAs from the indicated cells lines were used for RPA with IRF-7C and GAPDH-specific probes. The images in the same box indicate that they are derived from the same gels. The identities of RNAs are shown.</p

Intramuscular nerve distribution in the FPB (right, deep side), drawing demonstrating the sFPB was supplied by the branch of the median nerve and the dFPB was supplied by the branch of the ulnar nerve (MNB, branch of the median nerve; UNB, branch of the ulnar nerve).

<p>The scale bar represents 4 mm.</p

Intramuscular nerve distribution in the ADM (right, deep side), drawing demonstrating the branching pattern of the branch of the ulnar nerve.

<p>The scale bar represents 4 mm.</p

Light microscopic views of muscle spindles in the OP.

<p>Transverse section of the OP showing 2 spindles (arrow) arranged side-by-side and forming a paired complex. Their outer capsules are fused but their inner contents remain separate distinct. Each spindle contains several intrafusal fibers (arrowheads). The scale bar represents 100 µm.</p

Intramuscular nerve distribution in the AP (right, deep side), drawing demonstrating the oAP and tAP were supplied by the branch of the ulnar nerve (UNB, branch of the ulnar nerve).

<p>The scale bar represents 4 mm.</p

LMP-1 stimulates the expression of IRF-7C.

<p>DG75 cells were transfected with pcDNA3, LMP-1, or LMP-DM expression plasmids. The transfected cells were isolated, and total RNAs were isolated and used for RPA with IRF-7C and GAPDH-specific probes. Yeast RNA was used as negative control. Specific protections of IRF-7C and GAPDH RNAs are indicated.</p

Intramuscular nerve distribution in the APB (right, deep side), drawing demonstrating several intramuscular branches arising from the branch of the recurrent nerve of median nerve (NT, nerve trunk).

<p>The scale bar represents 4 mm.</p

K-RTA and EBV-Z interact with each other in dually infected cells.

<p>A. Co-localization of K-RTA and EBV-Z in dually infected cells. BC1 (KSHV+/EBV+) cells were treated with TPA (10 ng/ml) first for one day and then butyrate (0.5 mM) for another day. Cells were fixed and stained with K-RTA (rabbit) and EBV-Z (mouse) antibodies. Cy5- and Cy2-labeled secondary antibodies were used to distinguish the signals from K-RTA and EBV-Z, respectively. DAPI was used to stain the nuclei. The colors were artificially mounted to facilitate viewing. Red, K-RTA; green, EBV-Z; blue, nuclei; (a)K-RTA signal only; (b) EBV-Z signal only; (c) nuclei only; (d) K-RTA, EBV-Z, and nucleus signals are mixed. The pictures of higher power are shown on the bottom. In Panels B and C, cell extracts from treated BC1 cells were immunoprecipitated (IP) with either anti-EBV-Z or normal mouse serum (Panel B). Cell lysates were also immunoprecipitated with anti-K-RTA or normal rabbit serum (Panel C). The immunoprecipitates were analyzed by Western blot using the indicated antibodies. The whole cell lysates of induced BC1 cells were used as positive controls in Panels B and C. In Panel D, whole cell lysates was used for western blot analyses. The identity of the respective proteins is denoted. n.s., non-specific. Molecular weight (MW) makers are shown on the left in kilo-Dalton (kDa).</p

IRF-7C protein is associated with type III latency.

<p>A. Schematic diagram of IRF-7C-specific epitope. The open bar represents open-reading frames (ORF). The expression plasmids were made as shown. The IRF-7C has unique 13 amino-acid (aa), represented by solid bar, at the C-terminus because the splicing changes the original ORF. The peptide was synthesized according to the last 15-aa sequence in IRF-7C and was used for antibody production. IRF-7CD is an expression plasmid that lacks the C-terminal 13-aa of IRF-7C. Specific epitope for IRF-7C is shown. The drawing is not on scale. B. Examination of IRF-7C-specific antibody. 293T cells were transfected with various expression plasmids as shown on the top. The expression levels of IRF-7C and GAPDH proteins were determined by Western blotting. Left panel: IRF-7C-specific antibody was used. Right panel: full-length IRF-7 antibody was used. C. IRF-7C protein is highly expressed in cells with EBV type III latency. Cell lysates from the indicated cells lines were separated by 12% SDS-PAGE. The expression levels of IRF-7C and tubulin proteins were determined by Western blotting. The images in the same box indicate that they are derived from the same membrane. The identities of proteins are shown.</p

EBV-Z inhibits KSHV lytic gene expression.

<p>A. EBV-Z inhibits KSHV lytic gene expression. BC3 (KSHV+/EBV−) cells were transfected with CD4 expressing plasmid along with EBV-Z or vector plasmids. The transfected cells were isolated and equally split into two wells: one well of the cells was treated with TPA for 24 hours. Cell lysates were used for western blot analysis. B. Schematic of EBV-Z functional domains and mutants. The activation domain, basic region (DNA binding domain), leucine zipper region (LZ), and a region of unknown structure at the C terminus (CT) are shown. The drawing is not on scale. In Panels C, D, and E, 293T (EBV−/KSHV−) cells were transfected with various expression plasmids as shown on the top. FLAG-EBV-Z, and its mutants were used. Cell extracts from these transfected cells were immunoprecipitated with either anti-FLAG (for EBV-Z) (Panel C) or anti-K-RTA (Panel D). The immunoprecipitates were analyzed by Western blot using the indicated antibodies. In Panel E, whole cell lysate was used for western blot analyses. The identity of the respective proteins is denoted. In Panels F, G, and H, 293T (EBV−/KSHV−) cells were used. Panel F, KSHV Pan-promoter reporter construct (Pan-luc) and CMV-β-gal expression plasmid were cotransfected with 400 ng of EBV-Z or its mutant expression plasmids, together with 0, 20, 50 ng of K-RTA expression plasmids respectively as shown on the top. In Panel G, KSHV K14-promoter reporter construct (K14A-luc) and CMV-β-gal expression plasmid were cotransfected with 100 ng of EBV-Z or its mutant expression plasmids, together with 0, 10, 20 ng of K-RTA expression plasmids respectively as shown on the top. Luciferase activity was normalized by β -galactosidase activity. The relative folds of activation of promoter constructs are shown with standard deviations. One representative of three independent experiments is shown. Panel H, cell lysates from Panel F were used for western blot analysis. The same membrane was stripped and reprobed with other antibodies. The identity of proteins is as shown.</p