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. 2021 Mar 26;296:100599. doi: 10.1016/j.jbc.2021.100599

Figure 5.

Figure 5

S1P suppresses β-TrCP expression by upregulating miR-135b.A, PASMCs were stimulated with 1000-nM S1P for 48 h; the protein level of β-TrCP was examined using Western blotting, β-actin served as a loading control (n = 4 each group). B, PASMCs were transfected with the miRNA-135b inhibitor or negative control inhibitor; the expression of miRNA-135b was detected using qRT-PCR, U6 served as a loading control (n = 3 each group). C, PASMCs were previously transfected with the miRNA-135b inhibitor or negative control inhibitor for 24 h and then treated with 1000-nM S1P for 48 h; mRNA level of β-TrCP was detected by qRT-PCR, and β-actin served as a control (n = 3 each group). D, PASMCs were previously transfected with miRNA-135b inhibitor or negative control inhibitor for 24 h and then treated with 1000-nM S1P for 48 h; the protein level of β-TrCP was detected using Western blotting, β-actin served as a loading control (n = 4 each group). ∗p < 0.05 versus control, #p < 0.05 versus negative control inhibitor, &p < 0.05 versus negative control inhibitor and S1P-cotreated cells. β-TrCP, β-transduction repeat-containing protein; PASMCs, pulmonary artery smooth muscle cells; qRT, quantitative real-time; S1P, sphingosine-1-phosphate.