Figure 7: miR-103 promotes inflammatory activation of ECs by targeting KLF4. | Nature Communications

Figure 7: miR-103 promotes inflammatory activation of ECs by targeting KLF4.

From: Endothelial Dicer promotes atherosclerosis and vascular inflammation by miRNA-103-mediated suppression of KLF4

Figure 7

(a,b) Enrichment of Klf4 and c-Myb transcripts in the miRNA-induced silencing complex (RISC) of mouse aortic ECs (a) and human aortic ECs (HAECs, b) treated with miR-103-mimics as determined by GW182-IP. The results are expressed as the fold enrichment of the mRNAs in GW182-IP samples compared with the input samples. Results of two independent experiments are shown. ND indicates not detected. (c) Immunoblot analyses of KLF4 protein expression in HAECs treated with LNA-inhibitors of miR-103, non-targeting control LNA-oligonucleotides or premade KLF4 mRNA (n=3–4 per group). The KLF4 protein levels were normalized to those of GAPDH. Full scans of western blots are provided in Supplementary Fig. 14. (d) KLF4 mRNA levels in HAECs treated with LNA-inhibitors of miR-103 or control LNA-oligonucleotides (n=4–5 per group). (e) MiR-103 expression after silencing KLF4 using siRNA (siKLF4) in HAECs. A non-targeting siRNA (siNTC) was used in the control group (n=4–5 per group). (f) The expression of CXCL1, CX3CL1 and CCL2 after silencing KLF4 (siKLF4) in HAECs treated with (right) or without (left) LNA-inhibitors of miR-103 (n=4–5 per group). siNTCs were used in the control group. (g,h) The effect of transfection with GFP mRNAs (Ctrl) or premade KLF4 mRNAs on miR-103 (g) and chemokine expression (h) in HAECs treated with (h, right) or without (g; h, left) miR-103-mimics (n=3–5 per group). (i) Expression of CXCL1, CX3CL1 and CCL2 in HAECs treated with LNA-oligonucleotides (KLF4-target site blockers; KLF4-TSBs) designed to inhibit the interaction between miR-103 and the 3′UTR of KLF4 (n=6 per group). Non-targeting LNA-oligonucleotides were used in the control group. (j) Flow chamber assays to determine monocyte adhesion to HAECs treated with KLF4-TSBs or non-targeting oligonucleotides (n=4 per group). The data are represented as the mean±s.e.m. of the indicated number (n) of repeats. *P<0.05, ** P<0.01 and *** P<0.001 by Student’s t-test (a,b,d–j) and one-way analysis of variance (c).

Back to article page