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. 2015 Dec 15;10(12):e0144840. doi: 10.1371/journal.pone.0144840

Fig 2. MyD88 short is a negative regulator of NTHi-induced inflammation in airway epithelial cells.

Fig 2

(A-B) BEAS-2B cells were transfected with a Myc-tagged MyD88s overexpression plasmid or empty vector and (A) MyD88 protein was visualized via western blot analysis using antibodies against Myc and (B) Relative quantity of human MyD88s mRNA was measured by real-time Q-PCR analysis. (C-D) Cells were transfected with the MyD88s overexpression plasmid alone or together with a NF-κB luciferase reporter construct followed by NTHi stimulation for 6 hours. (C) Relative NF-κB promoter activity was measured. (D) Relative quantities of human IL-6 or human IL-1β mRNA were measured by real-time Q-PCR analysis. (E) Cells were transfected with MyD88s siRNA or control siRNA and relative quantity of MyD88s mRNA was measured by real-time Q-PCR analysis. (F-G) Cells were transfected with the MyD88s siRNA or control siRNA alone or together with a NF-κB luciferase reporter construct followed by NTHi stimulation for 6 hours. (F) Relative NF-κB promoter activity was measured. (G) Relative quantities of IL-6 or IL-1β mRNA were measured by real-time Q-PCR analysis. Data are mean ± SD (n = 3). *p<0.05. Statistical analysis was performed using Student’s t-test. Data are representative of three or more independent experiments.