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. 2017 Feb 16;292(17):6855–6868. doi: 10.1074/jbc.M117.775205

Figure 2.

Figure 2.

Determination of NO production and iNOS expression in M. smegmatis pSMT3-, M. smegmatis esxL-, M. tuberculosis H37Rv-, and MtbΔesxL-infected cells. A, RAW 264.7 cells were infected with M. smegmatis (Msm) pSMT3 and recombinant M. smegmatis esxL for 2 h. The production of NO was quantified using Griess reagent 24 and 48 h after infection. B, transcript levels of iNOS in M. smegmatis pSMT3- and M. smegmatis esxL-infected macrophages were determined by qRT-PCR 24 h post-infection. GAPDH was taken as internal control. iNOS expression was checked by Western blotting (C) and fluorescence-microscopic analysis (D) using iNOS-specific antibody in M. smegmatis pSMT3- and M. smegmatis esxL-infected macrophages. E, THP-1 cells were infected with M. tuberculosis (Mtb) and MtbΔesxL mutant for 2 h. The level of iNOS was checked by Western blotting 24 h post-infection. The experiments were performed in triplicate (n = 3). Results are shown as mean ± S.D. (error bars). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001.