Figure 4
(A) HRMECs were transfected with si-SOCS6 and/or miR-19b inhibitor. Transfection efficiency was detected by qRT-PCR. (B) HRMECs were transfected with miR-19b inhibitor, si-SOCS6, miR-19b inhibitor and si-SOCS6, si-SOCS6 and treated with WP1066 or si-NC. Cell viability was tested by the MTT assay. (C) The protein levels of the proliferation-related molecules CNA, Cyclin D1 and Cyclin E1 in HG-induced hRMECs were determined by Western blotting assay. (D) The activity of caspase-3/7 in HG-induced hRMECs was measured by commercial kits. (E) Cell apoptosis in HG-treated hRMECs was evaluated by flow cytometry analysis. (F) The levels of the apoptosis-related proteins cleaved caspase-3, Bcl-2 and Bax in HG-induced hRMECs were measured by Western blotting. (G and H) The secretion of inflammatory factors (TNF-α, IL-6 and IL-1β) in HG-induced hRMECs was measured by qRT-PCR and ELISA. (I) The protein levels of the signalling pathway components SOCS6, phosphorylated JAK2 (p-JAK2), total JAK2 (t-JAK2), phosphorylated STAT3 (p-STAT3) and total STAT3 (t-STAT3) were detected by Western blotting; n=3, *P<0.05, **P<0.01, ***P<0.001.
MiR-19b regulates apoptosis and inflammation by targeting SOCS6 via the JAK2/STAT3 signalling pathway in HG-induced hRMECs

(A) HRMECs were transfected with si-SOCS6 and/or miR-19b inhibitor. Transfection efficiency was detected by qRT-PCR. (B) HRMECs were transfected with miR-19b inhibitor, si-SOCS6, miR-19b inhibitor and si-SOCS6, si-SOCS6 and treated with WP1066 or si-NC. Cell viability was tested by the MTT assay. (C) The protein levels of the proliferation-related molecules CNA, Cyclin D1 and Cyclin E1 in HG-induced hRMECs were determined by Western blotting assay. (D) The activity of caspase-3/7 in HG-induced hRMECs was measured by commercial kits. (E) Cell apoptosis in HG-treated hRMECs was evaluated by flow cytometry analysis. (F) The levels of the apoptosis-related proteins cleaved caspase-3, Bcl-2 and Bax in HG-induced hRMECs were measured by Western blotting. (G and H) The secretion of inflammatory factors (TNF-α, IL-6 and IL-1β) in HG-induced hRMECs was measured by qRT-PCR and ELISA. (I) The protein levels of the signalling pathway components SOCS6, phosphorylated JAK2 (p-JAK2), total JAK2 (t-JAK2), phosphorylated STAT3 (p-STAT3) and total STAT3 (t-STAT3) were detected by Western blotting; n=3, *P<0.05, **P<0.01, ***P<0.001.

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