Figure 5
The effects of CCM111 on NF-κB and STAT3 pathway activities in murine RAW264.7 macrophages. (a) The cells were treated with LPS (100 ng/ml) and CCM111 (0, 80, 160 or 240 μg/ml) for 4 hours. (b) The cells were treated with 100 ng/mL LPS alone or with different concentrations of CCM111 (0, 160 and 240 μg/ml) for 4 h. (c) The cells were treated with IL-6 (10 ng/ml) and CCM111 (0, 60, 120 or 160 μg/ml) for 1 hour. (d) The cells were treated with IL-6 (10 ng/ml) alone or in combination with CCM111 (160 μg/ml) for 0.5, 1, and 2 hours. (e) The cells were treated with LPS (500 ng/ml) and CCM111 (0, 80, 160 or 160 μg/ml) for 4 hours. (f) The cells were treated with LPS (500 ng/ml) alone or in combination with CCM111 (160 μg/ml) for 2, 4, and 6 hours. (g) The cells were treated with IL-6 (10 ng/ml) alone or in combination with CCM111 (0, 60, 120, 160 μg/ml) for 0.5 hours. (a–g) After treatment, the cell lysates were analyzed by immunoblotting. (a) The protein levels of IκBα and phospho-IκBα were detected, and GAPDH was used as the internal control. (b) The protein levels of p65, IκBα expression were detected. β-actin was used as the cytosol internal control, and Histone H3 was used as the nuclear internal control. (c–f) The protein levels of STAT3 and phospho-STAT3 were detected, and β-actin was used as the internal control. (g) The protein level of phospho-Tyk2 was detected, and GAPDH was used as the internal control. The full length blots images are presented in Supplementary Fig. 3. The results were obtained from three independent replicates.
