Journal: Journal of Virology

Article Title: CD317 functions as a key antiviral factor in human herpesvirus 6 (HHV-6) infection

doi: 10.1128/jvi.00841-25

Figure Lengend Snippet: HHV-6 infection induces CD317 expression in host cells. ( A ) Effect of IFN-β stimulation on CD317 expression. Western blot analysis of CD317 expression in untreated and IFN-β-treated MT4 cells at 24 and 48 hours post-treatment. β-actin was used as an internal control. ( B ) IFN-β expression in control and HHV-6-infected cells. HHV-6-infected MT4 cells were harvested at 72 hpi, and IFN-β mRNA expression was quantified by RT-qPCR, normalized to β-actin expression. ( C ) Expression of CD317 in mock and HHV-6-infected T cells. MT4 cells were infected with HHV-6 or mock-treated, and CD317 levels were measured by Western blotting at 6, 12, 24, 48, and 72 hours post-infection. Vero cells served as a negative control, and HeLa cells were used as a positive control.

Article Snippet: Mouse monoclonal antibodies against β-actin and HRP-conjugated goat anti-mouse IgG (H + L) were obtained from Proteintech.

Techniques: Infection, Expressing, Western Blot, Control, Quantitative RT-PCR, Negative Control, Positive Control

Journal: Journal of Virology

Article Title: CD317 functions as a key antiviral factor in human herpesvirus 6 (HHV-6) infection

doi: 10.1128/jvi.00841-25

Figure Lengend Snippet: CD317 inhibits HHV-6 infection. ( A ) Expression of HHV-6 IE1 upon IFN-I stimulation. MT4 cells were stimulated with IFN-I for 24 and 48 hours, followed by HHV-6 infection for 24 hpi. Western blotting was performed to detect the expression of the HHV-6 IE1 protein in control and experimental groups. β-actin served as the internal control. ( B ) Endogenous CD317 expression analysis in host cells. Western blotting was used to detect endogenous CD317 in HHV-6-susceptible T cell lines (JJhan, HSB-2, MT4, and Molt3) and CBMCs. HeLa cells were used as a positive control. ( C ) Knockdown efficiency of CD317-targeting shRNAs. MT4 cells were transduced with the lentiviruses expressing control- or CD317-targeting shRNAs for 48 hours. Western blotting was used to assess CD317 expression, with β-actin as an internal control. ( D, E ) Expression of HHV-6 IE1 upon IFN-I stimulation after CD317 knockdown. MT4 cells were transduced with lentivirus for CD317 knockdown and stimulated with IFN-I for 24 and 48 hours. HHV-6 infection was performed after stimulation, and the expression of IE1 was analyzed by Western blotting. β-actin was used as an internal control.

Article Snippet: Mouse monoclonal antibodies against β-actin and HRP-conjugated goat anti-mouse IgG (H + L) were obtained from Proteintech.

Techniques: Infection, Expressing, Western Blot, Control, Positive Control, Knockdown, Transduction

Journal: Journal of Virology

Article Title: CD317 functions as a key antiviral factor in human herpesvirus 6 (HHV-6) infection

doi: 10.1128/jvi.00841-25

Figure Lengend Snippet: Overexpression of CD317 reduces HHV-6 infection. ( A ) Expression efficiency analysis of CD317. After transducing MT4 cells with control or CD317-expressing lentiviruses for 48 hours, CD317 expression was assessed by Western blotting, using β-actin as an internal control. ( B ) Effect of CD317 overexpression on HHV-6 attachment. MT4 cells overexpressing CD317 or the control cells were incubated with HHV-6 at 4°C for 1 hour, and infection was assessed by qPCR of HHV-6 genome. ( C ) Effect of CD317 overexpression on HHV-6 invasion. After attachment, cells were incubated at 37°C for 2 hours, and infection was analyzed by qPCR. ( D ) CD317 overexpression reduces HHV-6 entry. MT4 cells overexpressing CD317 or the control cells were infected with HHV-6, and expression of HHV-6 IE1 was detected by Western blot at 24 hpi. ( E and F ) CD317 overexpression decreases HHV-6 progeny production. MT4 cells were transduced with the control or CD317-expressing lentiviruses, infected with HHV-6 for 72 hours, and CV or SV levels were quantified by qPCR. Data are presented as fold change over control. Results are mean ± SD of three independent experiments. ns, P > 0.5; **** P < 0.01. ( G ) HHV-6 entry analysis using the viruses from CD317-overexpressing and control cells. The viruses from CD317-overexpressing cells and control cells were quantified and were used to infect MT4 cells. Intracellular HHV-6 DNA in target cells was detected via qPCR. Data are presented as fold change over the control. Results are the mean ± SD of three independent experiments. ns, P > 0.5; **** P < 0.01.

Article Snippet: Mouse monoclonal antibodies against β-actin and HRP-conjugated goat anti-mouse IgG (H + L) were obtained from Proteintech.

Techniques: Over Expression, Infection, Expressing, Control, Western Blot, Incubation, Transduction

Journal: Journal of Virology

Article Title: CD317 functions as a key antiviral factor in human herpesvirus 6 (HHV-6) infection

doi: 10.1128/jvi.00841-25

Figure Lengend Snippet: Knockdown of CD317 alone does not significantly affect HHV-6 infection. ( A ) Effect of CD317 knockdown on HHV-6 entry. MT4 cells were transduced with the control or shRNA-CD317 lentiviruses for 48 hours, infected with HHV-6 for 24 hours, and analyzed by Western blotting for IE1 expression. β-actin was used as an internal control. ( B and C ) Effect of CD317 knockdown on HHV-6 progeny production. MT4 cells transduced with control or shRNA-CD317 lentiviruses were infected with HHV-6 for 72 hours, and CV or SV were quantified by qPCR. Data are presented as fold change over control using the 2 -ΔΔCT method. Results are mean ± SD of three independent experiments performed in triplicate. ns indicates P > 0.5.

Article Snippet: Mouse monoclonal antibodies against β-actin and HRP-conjugated goat anti-mouse IgG (H + L) were obtained from Proteintech.

Techniques: Knockdown, Infection, Transduction, Control, shRNA, Western Blot, Expressing

Journal: Journal of Virology

Article Title: CD317 functions as a key antiviral factor in human herpesvirus 6 (HHV-6) infection

doi: 10.1128/jvi.00841-25

Figure Lengend Snippet: HHV-6 virions with low abundance of CD317 demonstrate enhanced efficiency of host cell entry. ( A ) CD317 is incorporated into HHV-6 virions. Purified HHV-6 virions were fixed, labeled with an anti-CD317 primary antibody, followed by a gold-conjugated secondary antibody, and then visualized using a transmission electron microscope. Scale bar: 25 nm. ( B ) Efficiency of CD317 knockdown in CBMCs. CBMCs were transduced with the lentiviruses for control- or CD317-shRNA expression for 48 hours, and CD317 expression was analyzed by Western blotting, with β-actin as an internal control. ( C ) CD317 detection in HHV-6 virions. HHV-6 was used to infect CD317-knockdown and control cells. After 72 hours, supernatants from these cells were collected, and HHV-6 virions were purified via ultracentrifugation. Western blotting was used to detect CD317 in virions, with gp96 and gH as controls. ( D ) Effect of CD317 in HHV-6 virions on the virus infection. An entry assay was performed on CBMCs using the viruses from CD317-knockdown and control cells. Intracellular HHV-6 DNA was detected via qPCR. Data are presented as fold change over the control. Results are the mean ± SD of three independent experiments. **** P < 0.01.

Article Snippet: Mouse monoclonal antibodies against β-actin and HRP-conjugated goat anti-mouse IgG (H + L) were obtained from Proteintech.

Techniques: Purification, Labeling, Transmission Assay, Microscopy, Knockdown, Transduction, Control, shRNA, Expressing, Western Blot, Virus, Infection

Journal: Veterinary Research

Article Title: Cytotoxicity induced by Aeromonas schubertii is orchestrated by a unique set of type III secretion system effectors

doi: 10.1186/s13567-025-01548-2

Figure Lengend Snippet: Proposed model of T3SS effector actions delivered by the API1 injectisome in A. schubertii. During interaction with host cells, A. schubertii translocates several T3SS effectors into the host cytosol via the API1 injectosome. AopL induces caspase-3/-7-independent necrosis, possibly by targeting the lysosomal V-ATPase, a prediction supported by its homology with VopQ from Vibrio parahaemolyticus . In contrast, AopU promotes caspase-3/-7-dependent apoptosis. These cytotoxic effects are counteracted by AopI, a pro-survival effector, which, based on its structural similarity to P. aeruginosa ExoY, is presumed to act as a nucleotidyl cyclase. Sequence homology and domain analysis also suggest that AopU, AopH, and AopO induce cell rounding and inhibit phagocytosis, while AopI and AopJ may interfere with host immune signaling pathways. The specific role of AopT remains uncertain.

Article Snippet: For loading control, membranes were re-probed with mouse monoclonal IgG against β-actin (dilution 1:10 000; Proteintech, Cat. No. 66009-1-Ig) and revealed with secondary HRP-anti-mouse IgG (Cytiva), as above.

Techniques: Sequencing, Protein-Protein interactions