Figure 1

Pro-inflammatory stimuli upregulate CD40 and reduce miR-424 and miR-503 expression in HUVECs. (a) CD40 mRNA expression in response to LPS (1 μg/ml) or TNFα (10ng/ml) in a time dependent manner. (b) CD40 protein expression in response to stimulation with either LPS (1 μg/ml for 24 h) or TNFα (10ng/ml for 16 h). (c) CD40 protein expression in HUVECs 72 h after AGO2 or DICER siRNA transfection. (d) Predicted target sequences of CD40 3′ UTRs targeted by miR-424 and miR-503 and mutated sequences (MUT#1, #2) for disrupting miR-424 and miR-503 recognition sequence. (e,f) Quantitative PCR showing expression of the mature and pri forms of miR-424 and miR-503 in response to LPS (1 μg/ml) or TNFα (10ng/ml) in a time dependent manner. (f) *P < 0.05, **P < 0.01, ***P < 0.001 compared to controls. Error bars, s.e.m. Cropped gel images are shown; uncut gels are included in the Supplementary Information.