Journal: Japanese journal of ophthalmology

Article Title: Infection of endotheliotropic human cytomegalovirus of trabecular meshwork cells.

doi: 10.1007/s10384-018-0618-1

Figure Lengend Snippet: Fig. 1 Induction of the mRNA of the genes of human cytomegalovirus (HCMV) in human trabecular meshwork cells (HTMC) after HCMV infection. HTMCs were infected with the Towne strain or the TB40/E strain of HCMV, and the induction of the mRNA of IE1 (a), pp65 (b), and glycopro- tein B (gB) (c) was determined by RT-real-time PCR. Viral genes, IE1, pp65, and gB, were sequentially induced after infection by the Towne strain or the TB40/E strain of HCMV. TB40/E induced significantly higher amounts of IE1, pp65, and gB. *P <0.05, **P <0.05; n = 8

Article Snippet: The transferred membranes were stained with antibodies for HCMV IE1(NEA-9221, Perkin Elmer), pp65 (CA003, EastCoast Bio), or UL44 (CH167, Santa Cruz) [5].

Techniques: Infection, Real-time Polymerase Chain Reaction

Journal: Japanese journal of ophthalmology

Article Title: Infection of endotheliotropic human cytomegalovirus of trabecular meshwork cells.

doi: 10.1007/s10384-018-0618-1

Figure Lengend Snippet: Fig. 6 Induction of viral gene proteins and remodeling of actin cytoskeleton after HCMV infection. HTMCs were infected with the TB40/E strain (a) or the Towne strain (b) of HCMV at MOI of 5 and immunostained for IE1, pp65, and glycoprotein B (Green). TB40/E infection induced significant remodeling of the actin cytoskeleton (c, pp65, green). Actin was stained using phalloidin (red). Nucleus was labelled by DAPI (blue). *Viral assembly compartment

Article Snippet: The transferred membranes were stained with antibodies for HCMV IE1(NEA-9221, Perkin Elmer), pp65 (CA003, EastCoast Bio), or UL44 (CH167, Santa Cruz) [5].

Techniques: Infection, Staining

Journal: Scientific Reports

Article Title: Human cytomegalovirus hijacks the autophagic machinery and LC3 homologs in order to optimize cytoplasmic envelopment of mature infectious particles

doi: 10.1038/s41598-019-41029-z

Figure Lengend Snippet: Roles of the autophagy machinery in different steps of the viral cycle. ( A ) Quantification of HCMV entry by immunofluorescence detection of the viral tegument protein pp65 2 h after contact with the virus (MOI 1). DAPI was used to stain nuclei. Percentages of pp65-positive cells correspond to viral entry into the indicated cell lines compared to control cells (deficient: Autophagy-deficient). ( B ) Quantification of HCMV genome replication 4 dpi at MOI 0.5 by real time PCR in the indicated cell lines, compared to control cells. ( C ) Expression of several viral proteins after HCMV infection of control and autophagy-deficient cell lines at MOI 0.5 for 1, 2 or 5 days post infection (dpi). Actin was used as a loading control. Full-length blots are presented in Fig. . ( D ) Indicated control and deficient cells were infected at MOI 0.5 with HCMV AD169 strain. Cell-associated viruses were collected 4 dpi and the amount of infectious virus was measured. Results represent the mean values of 3 independent experiments. ns: non-significant (Student’s t test).

Article Snippet: To detect HCMV-infected cells, we used mouse monoclonal antibodies directed against the viral proteins IE1 and IE2 (clone E13; Biomerieux, 11–003), tegument proteins pp65 (Novus, B051M) and pp28 (Santa Cruz, clone CH19, sc-69749).

Techniques: Immunofluorescence, Virus, Staining, Control, Real-time Polymerase Chain Reaction, Expressing, Infection

Journal: PLoS ONE

Article Title: Phenotypic characterization and anticancer capacity of CD8+ cytokine-induced killer cells after antigen-induced expansion

doi: 10.1371/journal.pone.0175704

Figure Lengend Snippet: Left: control group with no stimulation and fluorescence-labeled antibody addition; Middle: CD28 antibody stimulation alone; Right: CD28 antibody and CMV-pp65 antigen costimulation.

Article Snippet: For TCR-induced degranulation assay, 6 x 10 5 CIK cells in 100 μl PBS were stimulated with 1 μg/ml CD28 antibody (BD Biosciences), 1 μg/ml CMV-pp65 antigen polypeptides (Miltenyi) and stained with 5μμl PE-CD107a antibody (Miltenyi).

Techniques: Control, Fluorescence, Labeling