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Figure 4 | Respiratory Research

Figure 4

From: Effects of cigarette smoke on degranulation and NO production by mast cells and epithelial cells

Figure 4

RT-PCR analysis of NOS expression.Panel A. Time course of the effect of 1% CSS upon A549 iNOS mRNA expression. Upper panel: Lane 1: control, Lane 2: 3 h, Lane 3: 6 h, Lane 4: 24 h, Lane 5: 24 h CSS exposure and then cells returned to normal media for further 24 h; bottom panel: GAPDH corresponding to each sample. PCR gels shown are representative of three separate experiments. There was a decrease in iNOS mRNA in A549 cells reaching undetectable levels at 6 h, which persisted to 24 hr, even after further incubation in normal culture media. Panel B. Time course of the effect of 1% CSS on NHTBE eNOS mRNA expression. Upper panel: Lane 1: control, Lane 2: 3 h, Lane 3: 6 h, Lane 4: 24 h, Lane 5: 24 h CSS and returned to normal media for further 24 h; bottom panel: GAPDH corresponding to each sample. PCR gels are representative of three separate experiments. Similar changes were seen in the mRNA expression of eNOS in the NHTBE line as in the A549 cells with decreased levels at 3 h which persisted throughout the study period. Panel C. Effect of 1% CSS and 100 μM nicotine on rat RBL-2H3 eNOS mRNA expression. Upper panel: Lane 1: control, Lane 2–4: CSS 3 h, 6 h, 24 h, Lanes 5–7: nicotine 3 h, 6 h, 24 h; bottom panel: GAPDH corresponding to each sample. PCR gels are representative of three separate experiments. After CSS, there was a decline in mRNA at 3 h which returned to baseline by 24 h, while exposure to nicotine showed a decrease at 6 and 24 h.

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