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. 2020 Nov 25;21(23):8925. doi: 10.3390/ijms21238925

Figure 5.

Figure 5

Mal/ERK1/2–dependent IFNβ expression results in Ip-10 upregulation. (A,B) WT, Mal −/− and MyD88−/− iBMDMs (A) and macrophages isolated from bone marrow of wild type mice (BMDM WT) and Mal deficient mice (BMDM Mal−/−) (B) were treated with R848 (100 nM) or LPS (100 ng/mL) for 4 h. Thereafter, total RNA was isolated, converted to first-strand cDNA, and used as a template for quantitative real-time RT-PCR as described under “Materials and Methods.” Quantitative real-time RT-PCR was used to assay the expression levels of Ip-10. Experiments were repeated at least three times and data are presented in relative expression units where Hprt was used to normalize all samples and non-treated cells were assigned an arbitrary value of 1. (C,D) WT and Mal−/− iBMDMs were pretreated with DMSO (control), FR180204 (2 µM) (C), or JSH-23 (10 µM) (D) for 30 min. Next, cells were treated with R848 (100 nM) or LPS (100 ng/mL) for 4 h. Thereafter, total RNA was isolated, converted to first-strand cDNA, and used as a template for quantitative real-time RT-PCR as described under “Materials and Methods.” Quantitative real-time RT-PCR was used to assay the expression levels of Ip-10. Experiments were repeated at least three times and data are presented in relative expression units where Hprt was used to normalize all samples and non-treated cells were assigned an arbitrary value of 1. * p ≤ 0.001; ** p ≤ 0.01; *** p ≤ 0.05.