Figure 1
Figure 1. PVM replicates in mouse bone marrow–derived eosinophils. Electron micrographs of mouse bone marrow–derived eosinophils (bmEos) documenting morphologic features typical of eosinophils, including the bilobed nucleus (N) and specific granules in the cytoplasm (sg). Magnification: (A) ×2500, bar = 2 μm; (B-C) ×5000 and ×12000, respectively; bar = 500 nm. (D) Q-RT-PCR detection of virus replication in bmEos. bmEos were inoculated with PVM (●) or heat-inactivated PVM (hiPVM; ○) at a multiplicity of infection (MOI) of 1; *P < .05. Data shown are representative of 2 experiments performed in triplicate. (E) PVM N protein is detected in infected bmEos at day 8 after inoculation. Total protein extracts from control, PVM-infected, or hiPVM-challenged bmEos were probed with anti-PVM N peptide antibody (αNPVM). Anti–human GAPDH (αGAPDH) antibody was used as a control for protein loading. (F) Interferon-γ–induced protein (IP-10/CXCL-10) released from bmEos on day 4 after inoculation in response to PVM infection, hiPVM-challenge, or control challenge. Results are representative of 2 experiments performed in triplicate, P values as indicated. (G) Infectious virions are released from PVM-infected bmEos. Supernatants from bmEos cultures at 2 or 8 days after inoculation were used to challenge cells of the RAW 264.7 cell line, which is highly permissive for PVM replication.14 Total RNA from the RAW 264.7 cells was harvested at 5 days after inoculation, and Q-RT-PCR was performed to determine virus copy number. Results are data combined from 3 experiments, each performed in triplicate, **P < .01. Data represent mean ± SEM.

PVM replicates in mouse bone marrow–derived eosinophils. Electron micrographs of mouse bone marrow–derived eosinophils (bmEos) documenting morphologic features typical of eosinophils, including the bilobed nucleus (N) and specific granules in the cytoplasm (sg). Magnification: (A) ×2500, bar = 2 μm; (B-C) ×5000 and ×12000, respectively; bar = 500 nm. (D) Q-RT-PCR detection of virus replication in bmEos. bmEos were inoculated with PVM (●) or heat-inactivated PVM (hiPVM; ○) at a multiplicity of infection (MOI) of 1; *P < .05. Data shown are representative of 2 experiments performed in triplicate. (E) PVM N protein is detected in infected bmEos at day 8 after inoculation. Total protein extracts from control, PVM-infected, or hiPVM-challenged bmEos were probed with anti-PVM N peptide antibody (αNPVM). Anti–human GAPDH (αGAPDH) antibody was used as a control for protein loading. (F) Interferon-γ–induced protein (IP-10/CXCL-10) released from bmEos on day 4 after inoculation in response to PVM infection, hiPVM-challenge, or control challenge. Results are representative of 2 experiments performed in triplicate, P values as indicated. (G) Infectious virions are released from PVM-infected bmEos. Supernatants from bmEos cultures at 2 or 8 days after inoculation were used to challenge cells of the RAW 264.7 cell line, which is highly permissive for PVM replication.14  Total RNA from the RAW 264.7 cells was harvested at 5 days after inoculation, and Q-RT-PCR was performed to determine virus copy number. Results are data combined from 3 experiments, each performed in triplicate, **P < .01. Data represent mean ± SEM.

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