Culturing HFFs and Vaccinia Virus

HFFs (a gift from Dr. Nicholas Wallace) were cultured in Dulbecco’s modified Eagle’s medium (Quality Biological) containing 10% fetal bovine serum (Peak Serum) and 2 mM L-glutamine (Quality Biological). Cells were incubated in a 5% CO₂ atmosphere at 37°C. VACV Western Reserve strain (ATCC VR-1354), vD9muD10mu with both decapping enzymes inactivated, and vΔA23 with intermediate transcription factor gene A23 deleted were kindly provided by Dr. Bernard Moss and were described previously (22 (link), 23 (link)). Virus titer was determined by plaque assay as described elsewhere (32 (link)).

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Dhungel P., Brahim Belhaouari D, & Yang Z. (2023). La-related protein 4 is enriched in vaccinia virus factories and is required for efficient viral replication in primary human fibroblasts. Microbiology Spectrum, 11(5), e01390-23.

Publication 2023

Dulbecco’s modified Eagle’s medium L-glutamine

Assay Atmosphere Biological Cells Cultured medium Decapping enzymes Eagle Fetal bovine serum Gene transcription factor Glutamine Moss Plaque Serum Strain

Corresponding Organization : Texas A&M Health Science Center

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Variable analysis

independent variables

VACV Western Reserve strain (ATCC VR-1354)
VD9muD10mu with both decapping enzymes inactivated
VΔA23 with intermediate transcription factor gene A23 deleted

dependent variables

Virus titer determined by plaque assay

control variables

Dulbecco's modified Eagle's medium (Quality Biological) containing 10% fetal bovine serum (Peak Serum) and 2 mM L-glutamine (Quality Biological)
Incubation in a 5% CO2 atmosphere at 37°C

controls

Positive control: Not specified
Negative control: Not specified

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