nih 3t3  (ATCC)

Journal: PLOS Pathogens

Article Title: Eph receptor tyrosine kinases are functional entry receptors for murine gammaherpesvirus 68

doi: 10.1371/journal.ppat.1013263

Figure Lengend Snippet: (A) Schematic of Eph receptor-dependent block of MHV68 infection of susceptible cell lines (BioRender.com/jdlijfo). (B) Normalized read counts of the 14 Eph receptor genes in NIH 3T3 cells (GEO dataset series GSE196318 ). (C) Dose-dependent inhibition of MHV68 infection by soluble murine EphA4-Fc and EphB3-Fc but not EphA6-Fc on NIH 3T3 murine fibroblasts. MHV68 ORF59-GFP was pre-incubated with murine EphA4-Fc, EphA6-Fc or EphB3-Fc. EphA2-Fc and PBS were used as controls. GFP expression was measured by flow cytometry. Infection is indicated as percentage of GFP + cells normalized to PBS controls. Mean of three independent experiments, error bars represent SD. (D-G) Cell type-dependent inhibition of MHV68 infection by soluble murine Eph proteins at 100 nM homodimerized protein. EphA2-Fc and PBS were used as controls. GFP expression as indicator of infection was measured by flow cytometry. Infection is shown as percentage of GFP + cells normalized to PBS controls. Mean and symbols represent three individual experiments, error bars represent SD. Statistical significance was evaluated by ordinary one-way ANOVA, corrected by Holm-Šídák’s multiple comparisons test. *: p-value < 0.05, ***: p-value < 0.001, ****: p-value < 0.0001.

Article Snippet: NIH 3T3 (RRID:CVCL_0594) and NIH 3T12 (ATCC CCL-164, Manassas, VA) murine fibroblasts were maintained in DMEM supplemented with 8% FCS, 100 IU/mL penicillin, 100 μg/mL streptomycin, and 2 mM L-glutamine (D8).

Techniques: Blocking Assay, Infection, Inhibition, Incubation, Expressing, Flow Cytometry

Journal: PLOS Pathogens

Article Title: Eph receptor tyrosine kinases are functional entry receptors for murine gammaherpesvirus 68

doi: 10.1371/journal.ppat.1013263

Figure Lengend Snippet: (A) Schematic of virus neutralization by gH/gL-targeting nAbs. C57BL/6 mice were infected with 1,000 PFU MHV68 WT by intranasal inoculation. Serum was collected at 28 dpi (BioRender.com/6j9fwj6). (B) gH/gL-specific IgG from naïve or MHV68-infected C57BL/6 at a serum dilution of 1:80 was measured by MHV68 gL-gH ELISA. Background corrected optical density at 450 nm is shown. (C) Serum of MHV68-infected C57BL/6 mice neutralizes MHV68 infection on NIH 3T3 cells. MHV68 ORF59-GFP was pre-incubated with mouse serum at the indicated dilution. GFP expression was measured by flow cytometry. Infection is indicated as percentage of GFP + cells normalized to naïve serum. (D-E) Serum neutralization of MHV68 infection on NIH 3T3 cells is mediated by gH/gL-targeting antibodies. Antibodies to gH ecto /gL or gH D-I /gL were depleted using soluble complexes pre-coupled to magnetic beads. Fc was used as control. Neutralization was analyzed as in (C) at a serum dilution of 1:80. Micrographs were taken at 16 hpi. Mean and symbols representing individual experiments are shown. Statistical significance was evaluated by ordinary two-way ANOVA corrected for multiple comparisons by Tukey’s multiple comparisons test. ***: p-value < 0.001, ****: p-value < 0.0001.

Article Snippet: NIH 3T3 (RRID:CVCL_0594) and NIH 3T12 (ATCC CCL-164, Manassas, VA) murine fibroblasts were maintained in DMEM supplemented with 8% FCS, 100 IU/mL penicillin, 100 μg/mL streptomycin, and 2 mM L-glutamine (D8).

Techniques: Virus, Neutralization, Infection, Enzyme-linked Immunosorbent Assay, Incubation, Expressing, Flow Cytometry, Magnetic Beads, Control