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

Figure 3

Pellino3 knockout causes decreased cytokine secretion and increased virus multiplication. (a) Total RNA was isolated from WT and Peli3^-/- BMDMs and converted to first-strand cDNA. This was used as a template for conventional PCR amplifying genes as indicated. Products were resolved as described under Materials and Methods. (b) Wild-type and Peli3^-/- BMDMs were infected with VSV at MOI 1.5. Cell culture supernatants were collected 16 h of postinfection. Concentration of type I IFN was measured by bioassay; level of IP-10 and TNF was measured by ELISA as described under Materials and Methods. (c) Wild-type and Peli3^-/- BMDM cells were infected with VSV at MOI 0.5. Cell supernatants were collected 24 h of postinfection to determine viral titer as described under Materials and Methods. 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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