Fig. 2.

EGR1 expression is induced following VEEV infection in primary astrocytes. A) Primary astrocytes were infected with VEEV TC-83 at an MOI of 1 and total RNA extracted at 3, 6, 9, and 18hpi. Gene expression was determined using TaqMan assays for EGR1. Fold changes were calculated relative to 18S ribosomal RNA and normalized to mock samples using the ΔΔCt method. B) Cells were infected and RNA extracted as above in panel A. Viral genomic copies were determined by RT-qPCR. Results are displayed as genomic copies in logarithmic scale. C) Astrocytes were mock-infected or infected with TC-83 (MOI 1 or 5) for 1 h. At 18hpi, cell lysates were collected using blue lysis buffer and analyzed by immunoblot. PVDF membranes were probed to determine levels of EGR1 and VEEV capsid. β-Actin was used as a loading control. D) EGR1 protein levels were normalized to β-Actin. Data are expressed as the Mean ± SD (n = 3). *p-value≤ 0.05, **p-value ≤0.01, ***p-value ≤ 0.001.