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. 2015 Mar 23;97(6):1111–1119. doi: 10.1189/jlb.3A1114-557R

Figure 3. M.tb-mediated induction of iNOS mRNA and protein in human macrophages.

Figure 3.

Day 12 MDMs were incubated with M.tb (MOI 5:1) for 2 h, washed 3 times, and repleted with 2% autologous serum for 3, 6, and 24 h. Monolayers were lysed with TRIzol for extraction of total RNA or lysed with TN-1 buffer for whole-cell lysate at indicated time-points. (A) qRT-PCR was used to determine iNOS mRNA levels. Data were normalized to the β-actin gene, and RCN was determined. Shown are cumulative data from 5 independent experiments (mean ± sem; *P < 0.05). (B) Equal amounts of cell lysates were used to determine iNOS protein levels by Western blotting by use of iNOS antibody or β-actin antibody as a loading control. Shown is a representative experiment of 3 independent experiments.