https://orcid.org/0000-0003-0592-9102
Figures & data
Figure 1. Increased expression of the longest ANRIL transcript NR_003529 (NR) in coronary arteries from CAD patients vs. non-CAD controls. Coronary arteries from 6 CAD patients and 6 age-matched non-CAD individuals were studied with real-time RT-PCR analysis. (A) Schematic diagram for the structure of ANRIL transcripts NR_003529 and DQ485454, and circANRIL with exons 5–7. The location of the CAD haplotype block is shown. (B-E) The relative abundance of different transcripts of ANRIL in CAD patients vs. non-CAD controls as represented by expression of (B) exons 12–13b for the detection of NR_003529, (C) exons 5–7 for the detection of both NR_003529 and DQ485454, (D) exons 7–5 for the detection of circANRIL7-5, and (E) exons 11–13 for the detection of DQ485454. (F) exons 18–19 for the detection of 3ʹ end of NR_003529 *p < 0.05 and **p < 0.01 vs. vector, N = 6.
Figure 2. Overexpression of NR_003529 in HCAEC and EA.hy926 cells increases monocyte adhesion and transendothelial migration. (A) Expression of the NR_003529 transcript of ANRIL in HCAECs, HUVECs, and EA.hy926 cells transiently transfected with an expression plasmid for NR_003529 vs. pcDNA vector as quantified by qRT-PCR. (B) Expression of the DQ485454 transcript of ANRIL in HCAECs, HUVECs, and EA.hy926 cells transiently transfected with an expression plasmid for NR_003529 vs. pcDNA vector as quantified by qRT-PCR. (C) Relative adhesion of monocytes to HCAECs, HUVECs, and EA.hy926 cells transiently transfected with an expression plasmid for NR_003529 vs. pcDNA vector. (D) Relative transmigration of monocytes across a layer of HCAECs, HUVECs, and EA.hy926 cells transiently transfected with an expression plasmid for NR_003529 vs. pcDNA vector. *p < 0.05 and **p < 0.01 vs. vector, N = 5.
Figure 3. Knockdown of NR_003529 in HCAEC, HUVECs and EA.hy926 cells reduces monocyte adhesion and transmigration. (A) Expression of the NR_003529 transcript of ANRIL in HCAECs, HUVECs, and EA.hy926 cells transiently transfected with NR_003529-specific siRNA (siNR) vs. negative control siRNA (siNC) as quantified by qRT-PCR. (B) Expression of the DQ485454 transcript of ANRIL in HCAECs, HUVECs, and EA.hy926 cells transiently transfected with NR_003529-specific siRNA (siNR) vs. negative control siRNA (siNC) as quantified by qRT-PCR. (C) Relative adhesion of monocytes to HCAECs, HUVECs, and EA.hy926 cells transiently transfected with NR_003529-specific siRNA (siNR) vs. negative control siRNA (siNC). (D) Relative transmigration of monocytes across a layer of HCAECs, HUVECs, and EA.hy926 cells transiently transfected with NR_003529-specific siRNA (siNR) vs. negative control siRNA (siNC). *p < 0.05 and **p < 0.01 vs. vector, N = 5.
Figure 4. Overexpression of NR_003529 together with knockdown of DQ485454 in HCAECs dramatically increases monocyte adhesion and transmigration. (A) Expression of the NR_003529 transcript of ANRIL in HCAECs transiently co-transfected with DQ485454-specific siRNAs (siDQ1, siDQ2) vs. negative control siRNA (siNC) and the overexpression plasmid for NR_003529 vs. pcDNA vector as quantified by qRT-PCR. (B) Expression of the DQ485454 transcript of ANRIL in HCAECs transiently co-transfected with DQ485454-specific siRNAs (siDQ1, siDQ2) vs. negative control siRNA (siNC) and the overexpression plasmid for NR_003529 vs. pcDNA vector as quantified by qRT-PCR. (C) Relative adhesion of monocytes to HCAECs transiently co-transfected with DQ485454-specific siRNAs (siDQ1, siDQ2) vs. negative control siRNA (siNC) and the overexpression plasmid for NR_003529 vs. pcDNA vector. (D) Relative transmigration of monocytes across a layer of HCAECs transiently co-transfected with DQ485454-specific siRNAs (siDQ1, siDQ2) vs. negative control siRNA (siNC) and the overexpression plasmid for NR_003529 vs. pcDNA vector. *p < 0.05 and **p < 0.01 vs. vector, N = 5.
Figure 5. Overexpression of DQ485454 together with knockdown of NR_003529 in HCAECs dramatically reduces monocyte adhesion and transmigration. (A) Expression of the NR_003529 transcript of ANRIL in HCAECs transiently co-transfected with NR_003529-specific siRNAs (siNR1, siNR2) vs. negative control siRNA (siNC) and the overexpression plasmid for DQ485454 vs. pcDNA vector control as quantified by qRT-PCR. (B) Expression of the DQ485454 transcript of ANRIL in HCAECs transiently co-transfected with NR_003529-specific siRNAs (siNR1, siNR2) vs. negative control siRNA (siNC) and the overexpression plasmid for DQ485454 vs. pcDNA vector as quantified by qRT-PCR. (C) Relative adhesion of monocytes to HCAECs transiently co-transfected with NR_003529-specific siRNAs (siNR1, siNR2) vs. negative control siRNA (siNC) and the overexpression plasmid for DQ485454 vs. pcDNA vector. (D) Relative transmigration of monocytes across a layer of HCAECs transiently co-transfected with NR_003529-specific siRNAs (siNR1, siNR2) vs. negative control siRNA (siNC) and the overexpression plasmid for DQ485454 vs. pcDNA vector. *p < 0.05 and **p < 0.01 vs. vector, N = 5.
Figure 6. Expressions of EZR, CXCL11, and TMEM106B are differentially regulated by different transcripts of ANRIL. (A) Relative mRNA expressions of seven downstream genes of ANRIL (CLIP1, EZR, LYVE1, TMEM100, TMEM106B, CXCL11, and ENC1) in HCAECs transfected with the overexpression plasmid for either DQ485454 or NR_003529 transcripts of ANRIL vs. pcDNA3.1 vector control as determined by qRT-PCR. (B) Relative mRNA expressions of the seven downstream genes of ANRIL in HCAECs transfected with the NR_003529-specific siRNA (siNR) vs. negative control siRNA (siNC) as determined by qRT-PCR. *p < 0.05 and **p < 0.01 vs. vector, N = 5.
