Fig. 8: Validation of miR-1 targets in endothelial cells. | Communications Biology

Fig. 8: Validation of miR-1 targets in endothelial cells.

From: Cigarette smoke induces angiogenic activation in the cancer field through dysregulation of an endothelial microRNA

Fig. 8

A HUVECs were transduced with v-miR-1 (or control vector). Ago-RIP was performed on cell lysates and the levels of each gene in the Ago pull immunoprecipitate/input lysate was measured and expressed as 2−∆∆Ct. (n = 4, *p < 0.03. B–E HPMEC were transfected with miR-1 mimic (miR-1) or control RNA (Ctrl) and exposed to 10% CSE for 24 h. mRNA/ reference gene levels were measured, normalized to Ctrl in PBS group, and expressed as 2−∆∆Ct. B NOTCH3 (n = 6, *p < 0.05), (C) HS3ST1 (n = 6, *p < 0.05), (D) SEMA4B (n = 6, *p < 0.025,**p = 0.0026 (E) TFAP4 (n = 6, p = NS). F, G HUVECs were transfected with NOTCH3 siRNA or scrambled control RNA (ctrl), exposed to 10% CSE for the indicated times. F Cell numbers were determined and normalized to values at time ‘0’ (n = 3 per group and time, *p < 0.03, **p < 0.02). G De novo DNA synthesis was determined using a BRDU ELISA colorimetric assay. Values were normalized to the baseline (time ‘0’) and presented as relative BRDU incorporation. (n = 8 per group and time, *p < 0.05, **p < 0.0002).

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