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. 2017 Mar 9;7:44141. doi: 10.1038/srep44141

Figure 3. NOS2 protein amount and NO production in BMDMs infected with La-WT and La-arg− L. (L.) amazonensis.

Figure 3

BMDMs (1 × 106) were infected with La-WT (red) or La-arg− (blue) L. (L.) amazonensis (MOI 5:1). After 4 and 24 h of infection, the protein levels of NOS2 (A), the frequency of cells producing NO (B) and the average of fluorescence intensity reflecting NO production (C) were determined by in-cell western blotting and flow cytometry analysis of DAF-FM staining. Each bar represents the average ± SEM of the values obtained in 3 independent experiments, n = 6. Statistical significance was determined based on two-tailed Student’s t test. *p < 0.05, compared to uninfected macrophages (0 hours of infection). **p < 0.05, La-WT compared to La-arg−. (D) Sequence alignment of miR-294 and miR-721 and its target site in the 3′UTR of Nos2 using the microRNA.org database. Mmu-miR-294/Nos2 Alignment: mirSVR score −0.3992, PhastCons: 0.5607; Mmu-miR-721/Nos2 Alignment: mirSVR score −0.1183, PhastCons: 0.6227. Target sites of conserved miRNAs with good mirSVR scores.