Ligase Pellino3 Regulates Macrophage Action and Survival in Response to VSV Infection in RIG-I-Dependent Path

Figure 4

VSV-dependent ERK1/2 activation and VSV resistance are regulated by Pellino3. Wild-type and Peli3^-/- BMDMs were pretreated with DMSO (Mock) or FR180204 (2.0 μM for 1 h (b) or 5 μM for 72 h (c, d)). Next, the cells were infected with VSV at MOI 1.5 (a, b, c, d). (a) Cell lysates were subjected to SDS-PAGE followed by Western blotting. Protein detection was performed using specific antibodies and appropriate secondary antibodies conjugated to the fluorescent dye in the infrared range. Visualization was performed using the Odyssey CLx Imaging System LI-COR. The results presented are representative of at least three independent experiments. (b) Cell supernatants were collected 16 h of postinfection. Type I IFN was measured by bioassay; level of IP-10 and TNF was measured by ELISA as described under Materials and Methods. (c) Cell viability was evaluated by alamarBlue test 72 h of postinfection. (d) Photographs were taken using 10x microscope objective. Results are representative of at least three independent experiments performed in triplicate (). Statistical analysis was carried out using the unpaired Student -test using GraphPad Prism 7.04. values of less than or equal to 0.05 were considered to indicate a statistically significant difference (, , and ).

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