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Figure 5 | BMC Microbiology

Figure 5

From: c-KIT signaling is targeted by pathogenic Yersiniato suppress the host immune response

Figure 5

c-KIT signaling is targeted by Yersinia T3SS to suppress pro-inflammatory immune response. (A) THP1 cells were pre-treated with 1 μM OSI-930 or 1 μM BI-78D3 for 18 h or untreated prior to infection with Y. enterocolitica (pYV+) and Y. enterocolitica (pYV-) at MOI 2. The RNA levels are presented as fold change versus untreated THP1. Data is shown from three independent infection experiments performed in duplicate. A ‘*” denotes that relative RNA levels were significantly different (p<0.05) in OSI930-treated cells compared to untreated or BI-78D3-treated cells. (B) THP-1 cells were transfected with 50 nM siRNA targeting c-KIT or control (si-CTL) and incubated for 48 h. RNA levels are presented relative to transcript levels in siRNA-treated versus untreated THP-1. Data is shown from two representative experiments. A ‘*” denotes that relative RNA levels were significantly different (p<0.05) in si-cKIT-treated cells compared to si-CTL-treated cells. (C) THP-1 cells were transfected with 50 nM siRNA against c-KIT (si-cKIT) or control (si-CTL) siRNA and incubated for 72 h prior to infection with Y. enterocolitica WA at MOI 2 for 1 h. Gene transcript levels are depicted as a relative ratio to uninfected siRNA-treated THP-1 cells. Data is shown from three independent experiments performed in duplicate. A ‘*” denotes that relative RNA levels of immune genes were significantly different (p<0.05) in si-cKIT-treated cells compared to si-CTL-treated cells. (D) THP-1 cells, untreated or pretreated with 1μM OSI-930 for 5 h, were infected with Y. enterocolitica WA at MOI 40 for 45 min. Cell nuclei were purified, labeled with mouse anti-NF-κB RelA, and analyzed by flow cytometry. (left panel) The mean channel fluorescence was used to determine the fold change of RelA in the nuclei of Yersinia-infected compared to untreated THP-1 cells (middle panel). The statistical data was derived from two independent experiments (right panel).

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