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. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Cell Microbiol. 2009 Sep 2;12(1):67–83. doi: 10.1111/j.1462-5822.2009.01379.x

Fig. 2. OmpA+ E. coli induces NO production by increasing the expression of iNOS in HBMEC.

Fig. 2

(A) Confluent monolayers of HBMEC in 24-well culture plates were treated with OmpA+ or OmpA− E. coli for varying periods as indicated. Some monolayers were treated with AG or L-NAME for 1 h prior to addition of the bacteria. NO production in terms of nitrite was determined by Greiss method. (B) HBMEC monolayers infected with OmpA+ or OmpA− E. coli were washed, total cell lysates prepared, and subjected to Western blotting with antibodies to iNOS, eNOS or β-actin. Total cell lysates from uninfected monolayers were used as control. The numbers indicate fold increase in intensities of the bands compared to control and were normalized to β-actin levels (C) Total RNA was isolated from the cells as treated in B and subjected to RT-PCR using primers for iNOS, eNOS, or GAPDH. These data in A represent mean ± SD from three separate experiments performed in triplicate. The production of NO by OmpA+ E. coli was significantly greater in comparison to the levels induced by OmpA− E. coli at all time points, *P<0.001 by Student’s t test.