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. 2016 Sep 28;6:34445. doi: 10.1038/srep34445

Figure 5. NTHi-induced MyD88 short expression is mediated by MKP-1 up-regulation via a cAMP-PKA-dependent mechanism.

Figure 5

(a,b) BEAS-2B cells were treated with 8-bromo-cAMP 1 hour prior to NTHi stimulation. Relative quantities of human MyD88s (a) and human MKP-1 (b) mRNA were measured by real-time QPCR analysis. (c,d) Cells were treated with forskolin 1 hour prior to NTHi stimulation. Relative quantities of human MyD88s (c) and human MKP-1 (d) mRNA were measured by real-time QPCR analysis. (e) Cells were treated with resveratrol followed by NTHi and cell lysate was used to determine cAMP levels. (f) Western blot analysis of MKP-1 expression. Cells were treated with forskolin for 1 hour and stimulated with NTHi for 90 minutes. (g) Cells were transfected with shMKP-1 or empty vector and then treated with forskolin followed by NTHi stimulation. Relative quantities of human MyD88s mRNA were measured by real-time QPCR analysis. (h,i) Cells were treated with H89 1 hour prior to NTHi stimulation. Relative quantities of human MyD88s (h) and human MKP-1 (i) mRNA were measured by real-time QPCR analysis. (j,k) Cells were treated with PKI 1 hour prior to NTHi stimulation. Relative quantities of human MyD88s (j) and human MKP-1 (k) mRNA were measured by real-time QPCR analysis. (l) Western blot analysis of MKP-1 protein expression. Cells were treated with PKI for 1 hour followed by NTHI stimulation for 90 mins. (m) Cells were transfected with shMKP-1 or empty vector and then treated with PKI followed by NTHi stimulation. Relative quantities of human MyD88s mRNA were measured by real-time QPCR analysis. Data are mean ± SD (n = 3). *p < 0.05. Statistical analysis was performed using Student’s t-test. Displayed immunoblots are cropped images from full-length blots, present in Supplementary Figure S3. Data are representative of three or more independent experiments.