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. 2020 Jun 5;9(6):491. doi: 10.3390/antiox9060491

Figure 3.

Figure 3

IL-6/JAK2/STAT3 cascades participated in CRAMP inhibited VSMC Phenotypic transformation. (A) VSMC were pretreated with CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) for 24 h. The activation of ERK1/2 and STAT3 was detected using immunoblotting with anti-p-ERK1/2, anti-STAT3 antibodies. Data of 3 independent experiments is presented as mean ± SEM. * p < 0.05, *** p < 0.001 compared with control, n = 3. (B) VSMC were pretreated with CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) for 24 h. IL-6 in culture medium was detected by enzyme-linked immunosorbent assay (ELISA). Data of 3 independent experiments is presented as mean ± SEM. *** p < 0.001 compared with control, n = 4. (C) VSMC were pretreated with IL-6 antibody (10 μg/mL) for 2 h and CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) for 24 h followed by immunostaining with p-JAK2, p-STAT3 and SM22α antibodies. (D) VSMC were pretreated with recombinant IL-6 at final concentration of 10 ng/mL and CRAMP (100 ng/mL) for 2 h and then stimulated with PDGF-BB (20 ng/mL) for 24 h followed by immunostaining with p-JAK2, p-STAT3 and SM22α antibodies. Data of 3 independent experiments is presented as mean ± SEM. ** p < 0.01, *** p < 0.001 compared with control, n = 3.