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. 2020 Feb 6;35:100954. doi: 10.1016/j.molmet.2020.01.018

Figure 7.

Figure 7

Inflammatory and insulin signaling in enteroendocrine STC-1 cells stimulated with pro-inflammatory conditioned media. A) STC-1 cells were pre-incubated or not for 2 h with the PTP1B inhibitor (20 μM) before the treatment with mCM-LPS for 5, 15, or 30 min. The dishes pretreated with the PTP1B inhibitor were used throughout the experiment. At the end of the culture time, cells were lysed and protein extracts were analyzed by western blot with the corresponding antibodies. Vinculin was used as a loading control. Right panel shows representative western blots. In the left panel, graphs show quantification of three independent experiments performed in duplicate. ∗∗p < 0.01 mCM-LPS plus PTP1B inhibitor vs mCM-LPS. B) PTP1B inhibition protected against the drop of insulin signaling in EECs stimulated with pro-inflammatory conditioned medium. STC-1 cells were pre-incubated with the PTP1B inhibitor (20 μM) before the treatment with mCM-LPS or mCM-C for 16 h. Then, insulin was added for an additional 15 min. Protein extracts were prepared, and the phosphorylation of the IR and AKT was analyzed by western blot with the antibodies against phospho-IR, phospho-AKT Ser473, phospho-AKT Thr308, total IR, and total AKT. An anti-p85-PI3K antibody was used as a loading control. Right panel shows representative western blots. In the left panel, graphs show quantification of three independent experiments. The different experimental conditions are indicated under the blots or graphs. Data are shown as mean ± SEM. + p < 0.05 mCM-LPS plus PTP1B inhibitor vs mCM-LPS.