Figure 3: APLNR regulates CXCR4 expression through miR-139-5p.
From: MicroRNA 139-5p coordinates APLNR-CXCR4 crosstalk during vascular maturation
(a,b) CXCR4 expression in response to shear stress exposure in HUVECs, with or without concurrent APLNR knockdown. mRNA (a) and protein levels (b) are shown. (c) CXCR4 expression in response to AGO2 knockdown. (d) CXCR4 expression in response to shear stress exposure in HUVECs, with or without concurrent AGO2 knockdown. (e) Determination of miR-139-5p levels in FACS sorted ECs from P5 retinas of Aplnr−/−, Apln−/− mice and respective littermates. n=3 retinas per genotype. (f) CXCR4 expression in HUVECs overexpressing miR-139-5p mimic or non-targeting control anti-miR. (g) Expression levels of miR-139-5p in response to shear stress, with or without concurrent APLNR knockdown. (h) CXCR4 expression in HUVECs in response to shear stress exposure, with or without concurrent miR-139-5p inhibition via anti-miR. (i) Sprouting assay using HUVEC covered beads transduced with miR-139-5p or control lentivirus in response to SDF-1α or VEGF 165. Scale bar, 175 μm. (j) Migration assay of HUVECs transduced with miR-139-5p or control lentivirus in response to SDF-1α. Scale bar, 200 μm. *P<0.05, **P≤0.01, ***P≤0.001, ###P≤0.001 against NC v/PBS, †††P≤0.001 against NC v/SDF-1α, t-test. Error bars represent s.e.m. n=3 experiments per condition.
