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. 2020 Dec 9;95(1):e01768-20. doi: 10.1128/JVI.01768-20

FIG 2.

FIG 2

Abl2 behaves as a gene of the cellular innate antiviral response independent of interferon beta signaling. (A) The expression levels of genes associated with the IFN-β response, coding for IFN-β and for two ISGs (IRF7 and OAS1B), along with the gene coding for Abl2, were measured by RT-qPCR in L929 cells either noninfected (NI) or at different times after NDV infection. Relative expression levels were determined with respect to three reference genes (Utp6c, Ppib, and Hprt1) and fold inductions were calculated with respect to noninfected (NI) cells. Results correspond to the average of n = 3 independent experiments for all conditions tested. Data are means ± SD. Statistical analysis was carried out only for Abl2 gene expression using a one-way ANOVA test with Dunnett correction with each NDV-infected condition compared to noninfected conditions. (B and C) Abl2 protein levels were analyzed by Western blotting in extracts from L929 cells noninfected (NI) or infected by NDV. (C) Quantification using ImageJ with n = 3 independent experiments for NDV infection at 8, 20, and 24 h p.i. and n = 10 for NI. Data are means ± SD. Significance was assessed by one-way ANOVA test with Dunnett correction and each NDV-infected condition compared to the noninfected condition. (D) The expression levels of genes associated with the IFN-β response, coding for IFN-β and for two ISGs (IRF7 and OAS1B), along with the gene coding for Abl2, were measured by RT-qPCR in L929 cells collected at 6 h p.i. with NDV, either untreated or treated with increasing amounts of anti-IFNAR antibody. Relative expression levels were determined with respect to three reference genes (Utp6c, Ppib, and Hprt1) and fold inductions were calculated with respect to noninfected (NI) cells considered 100%. Results correspond to the average of n = 4 independent experiments for 0 μg; n = 2 for 0.5 μg; and n = 3 for 1.5 and 5 μg of anti-IFNAR antibody. Data are means ± SD. Significance was assessed by one-way ANOVA test with Bonferroni correction for each gene separately. P values: ***, <0.001; **, <0.01; and *, <0.05.