Skip to main content
. 2019 Aug 9;21(12):e13090. doi: 10.1111/cmi.13090

Figure 4.

Figure 4

Mtb FadD13 promotes the production of proinflammatory cytokines in macrophages through eEF1A1. (a) Western blot results showing H37Rv upregulated the expression of eEF1A1 in macrophage. The differentiated THP‐1 macrophages were infected at a MOI of 10 with H37Ra and H37Rv, respectively, followed by protein extraction and western blot. Blank: uninfected THP‐1 macrophages; H37Ra: H37Ra infected THP‐1 macrophages; H37Rv: H37Rv infected THP‐1 macrophages. The results presented as the one replicate from two independent experiments. (b) Western blot showing ΔfadD13 mutant decreased the expression of eEF1A1 in the macrophages. The differentiated THP‐1 macrophages were infected at a MOI of 10 with ΔfadD13 mutant and H37Rv respectively, followed by protein extraction and western blot. Blank: uninfected THP‐1 macrophages; ΔfadD13:ΔfadD13 infected THP‐1 macrophages; H37Rv: H37Rv infected THP‐1 macrophages. The results presented as the one replicate from two independent experiments. (c) Western blot result showing FadD13 upregulated the expression of eEF1A1 in overexpressed HEK293T cells. HEK 293T cells were transfected with the vectors encoding FadD13‐GFP or GFP alone respectively for 24 hr, followed by protein extraction and western blot. The results presented as the one replicate from two independent experiments. (d, e) ELISA results showing the production of proinflammatory cytokines (IL‐1β, IL‐18, and IL‐6) decreased in the eEF1A1 knockdown macrophages. NC: THP‐1 macrophages transfected with control siRNA. siRNA eEF1A1: THP‐1 macrophages transfected with eEF1A1 specific siRNA. The differentiated THP‐1 macrophages were infected at a MOI of 10 with FadD13 overexpressed M. smegmatis (d) or H37Rv (e) respectively. The results presented as the one replicate from three independent experiments. * p < .05 and ** p < .01 (two‐tailed unpaired t test)