Fig. 6: JunB overexpression or IFN-γ has little effect on NCOR2-013-mediated inhibition of inflammatory response in macrophages, while LPS has partial effect.
From: Talaromyces marneffei suppresses macrophage inflammation by regulating host alternative splicing
a JunB overexpression in NCOR2-013 overexpressing THP-1 macrophages was confirmed by RT-qPCR and WB (n = 4 biological replicates, data are presented as mean values ± SD). b, c The JunB overexpressing THP-1 macrophages and control cells were infected with T. marneffei conidia (MOI = 10) for 24 h. The expression of TNF-α and IL-1β was detected by RT-qPCR (b) (n = 3 biological replicates, data are presented as mean values ± SD). T. marneffei CFUs were detected in JunB overexpressing THP-1 macrophages and control cells (c). Four gradient serial dilutions (100, 10−1, 10−2, 10−3) were performed (n = 9 biological replicates, data are presented as mean values ± SD). d, e The NCOR2-013 overexpressing THP-1 macrophages and control cells were stimulated with IFN-γ (100 ng/mL) for 24 h, and then infected with T. marneffei conidia (MOI = 10) for 24 h. The expression of TNF-α and IL-1β was detected by RT-qPCR (d) (n = 3 biological replicates, data are presented as mean values ± SD). T. marneffei CFUs were detected in IFN-γ-stimulated NCOR2-013 overexpressing THP-1 macrophages and control cells (e). Four gradient serial dilutions (100, 10−1, 10−2, 10−3) were performed (n = 8 biological replicates, data are presented as mean values ± SD). f, g The NCOR2-013 overexpressing THP-1 macrophages and control cells were stimulated with LPS (500 ng/mL) for 24 h, and then were infected with T. marneffei conidia (MOI = 10) for 24 h and 48 h. The expression of TNF-α and IL-1β was detected by RT-qPCR (f) (n = 3 biological replicates, data are presented as mean values ± SD). T. marneffei CFUs were detected in LPS-stimulated NCOR2-013 overexpressing THP-1 macrophages and control cells (g). Four gradient serial dilutions (100, 10−1, 10−2, 10−3) were performed (n = 9 biological replicates, data are presented as mean values ± SD). All data are shown as mean ± SD from three independent experiments. WB were shown were the representative blot. Two-tailed Student’s t test was used to determine significance, denoted by *(P < 0.05), **(P < 0.01), and ns (not significant). Supplementary material is available (Supplementary Figs. 3, 6; Supplementary Data 1). MOI multiplicity of infection, NC negative control, NCOR2-013 OE NCRO2-013 overexpression, JunB OE JunB overexpression, Tm T. marneffei.
