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. 2021 Sep 2;17(9):e1009888. doi: 10.1371/journal.ppat.1009888

Fig 4. TBK1 is required for type I IFN induction in response to R. equi infection in primary murine macrophages.

Fig 4

(A) Immunoblot of pSTAT1, total STAT1, pIRF3 and total IRF3 in RAW 264.7 macrophages infected or not (0) with R. equi for 2, 4 or 6h or transfected with ISD for 4h. ACTIN was used as a loading control. (B) RT-qPCR of Ifnb in control (Tbk1+/+Tnfr-/-) and KO (Tbk1-/-Tnfr-/-) BMDMs transfected with ISD for 4h. (C) RT-qPCR of Il1b in TBK1 BMDMs uninfected or infected with R. equi for the indicated times. (D) As in (C) but Ifnb, Isg15 and Ifit1. (E) RT-qPCR of Isg15, Ifit1 and Mx1 in Myd88 KD macrophages infected with R. equi for the indicated times. All RT-qPCRs are representative of at least 2 independent experiments and are the mean of 3 replicates ± SD, n = 3. Immunoblots are representative of at least 3 independent experiments. Statistical significance was determined using Students’ t-test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, n.s. = not significant.