Figure 4: KSHV miRNAs inhibit IFN-αA-dependent STAT3 activation.
From: Viral microRNAs Target a Gene Network, Inhibit STAT Activation, and Suppress Interferon Responses
(A) HUVECs were transfected with KSHV miRNAs (miR-K6-5, miR-K8 or miR-K9*) or control miRNAs (miR-Neg). At 48 hpt, transfected cells were mock-treated or treated with IFN-αA for 20 minutes. Equal amounts of protein were probed for pY705-STAT3, total STAT3 and actin (top) or pY701-STAT1 and actin (bottom). (B) Relative levels of various proteins from 4A were normalized to actin, compared to the negative control miRNA (Neg), and plotted. (C) HUVECs were transfected with miR-Neg and treated with IFN-αA for 4 hours and mRNA levels of the indicated interferon-stimulated genes (ISGs) were measured by qRT-PCR. D. HUVECs were transfected with miRNAs as in (A) and treated with IFN-αA for 4 h and qRT-PCR was performed to measure levels of the indicated ISGs. Expression changes in ISGs were normalized to miR-Neg+ IFN-αA condition and plotted. Statistical significance relative to corresponding control RNAs with at least three biological replicates and p < 0.05 is denoted by *p < 0.001 by and **p < 0.0001 by. ***Full-length blots are in Figure S3.
