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

Figure 2

Immune response in BMDMs after VSV infection is RIG-I-dependent. (a) Wild-type and Mavs^-/- BMDM cells were infected with VSV at MOI 1.5. Cell culture supernatants were collected 16 h of postinfection. Type I IFN concentration was measured by bioassay; level of IP-10 and TNF was measured by ELISA as described under Materials and Methods. (b) A549-Dual, A549-Dual KO-MDA-5, and A549 KO-RIG-I cells were infected with VSV at MOI 1.5. After 16 h of the interferon regulatory factor (IRF) pathway was determined by monitoring activity of Lucia luciferase as described under Materials and Methods. Results are representative of at least three independent experiments performed in triplicate (). (c)WT BMDMs were infected with VSV at MOI 1.5 and incubated for 4, 8, and 16 h. Therefore, total RNA was isolated, converted to first-strand cDNA, and used as a template for quantitative real-time PCR as described under Materials and Methods. Quantitative real-time PCR was used to assay the expression levels of Peli3. Experiments were repeated at least three times, and data are presented in relative expression units where Hprt was used to normalize all samples and untreated cells were assigned an arbitrary value of 1. 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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