Fig. 2.

Inhibition of ERK1/2 phosphorylation reduces TC-83 replication and leads to lowered virus protein expression. (A) U87MG cells were treated with Ag-126 for 24 h, and the Cell-Titer-Glo assay was conducted. Luminescence was measured and the values indicated as luminescence units in the Y-axis. The concentrations of the drug that were used in this experiment are indicated in the X-axis. DMSO alone was used as a negative control, at 0.1% concentration. (B) U87MGs were pretreated Ag-126 for 2 h. Cells were infected with TC-83 for one hour (MOI:0.1). The conditioned media was then added back onto the cells and 24 hpi the supernatants were collected. The graph portrays the results of three independent experiments. (C) Cells were treated with 10 μM Ag-126 or DMSO alone and infected at either an MOI of 0.1, 0.5, or 1. Conditioned media was added back to the cells after the one hour infection period, and left until supernatants were collected at 24 hpi. (D) Whole cell lysates were obtained from U87MG cells after infection with TC-83 (MOI:1). Cells were infected in the presence or absence of Ag-126 treatment (10 μM). Infected cell lysates were collected at 8 hpi and 24 hpi. Amounts of β-actin were used as a loading control. Data in the bar graph is representative of duplicate experiments at the 24 hpi time point. (E) Cells were infected with TC-83 (MOI:1) and either untreated or treated with Ag-126. Total protein lysates were obtained at the indicated time points and analyzed by Western blot for phosphorylation status of ERK1/2.