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. 2020 Jun 24;11:1300. doi: 10.3389/fimmu.2020.01300

Figure 2.

Figure 2

SHH agonist and antagonist regulate JNK signaling. (A–E) Effects of SHH agonist (SAG) and antagonist (cyclopamine) on the phosphorylation of JNK and c-Jun were evaluated by Western Blot. FLSs were stimulated with SAG (1 μM) at different time points (5, 15, 30, 60, 120 min) and the levels of phospho-JNK and phospho-c-Jun were determined by Western Blot (n = 4) (A,C). To detect the effect of SHH antagonist on JNK signaling, FLSs were treated with cyclopamine (10 μM) for 24 and 48 h before the levels of phospho-JNK (B) and phospho-c-Jun (D) were detected, respectively (n = 5). The expression of phospho-c-Jun was evaluated after FLSs were treated with SAG in the presence of JNK inhibitor SP600125 for 15 min (n = 5) (E). Results were normalized to the levels of total JNK or c-Jun. FLSs in control group were treated with vehicle (DMSO supplemented in DMEM with 10%FBS). The results are shown as the mean ± S.D. Data were analyzed using ANOVA with Dunnett's post hoc test (A,C,E) or independent sample Student's t-test (B,D). *P < 0.05 vs. control group. **P < 0.01 vs. control group. ***P < 0.001 vs. control group. ns, non-significant; CON, control group; CYC, cyclopamine; SP, SP600125.