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. 2024 Apr 2;19:3123–3142. doi: 10.2147/IJN.S437714

Figure 4.

Figure 4

Suppression of the upregulated expression of collagen under laminar shear stress increased the uptake of VSOPs. (A) Gene expression of COL1A2, COL5A1, and COL5A2 in S- and LSS-cultivated HUVECs measured by RT-qPCR. Gene expression was normalized to the housekeeping gene ACTB. *P < 0.05 and **P < 0.01 LSS vs S by a ratio paired t-test. n = 4. (B) COL1A2 and (C) COL5A2 protein levels in untreated S- and LSS-cultivated HUVECs detected by Western blotting. Amido black staining shows equal loading. (D-G) Relative mRNA gene expression (measured by qRT-PCR) and VSOP particle uptake by LSS-cultivated HUVECs treated with (D and E) HF or (F and G) siRNAs against COL5A1/COL5A2. (E) and (G) Prussian blue stained VSOPs are shown as blue clusters around and within formaldehyde fixed cells. Particle uptake was measured by the percentage of Prussian blue stained area per ROI. For each experiment, 6 ROI per treatment were taken. Scale bar = 20 μm. (D and F) For gene expression analysis, relative target gene fold change (shown as 2-ΔΔC) was calculated using the comparative ΔΔCt analysis. For VSOP uptake n = 5 (HF) or n = 4 (siCOLV). Target Gene expression was normalized to the housekeeping gene ACTB for HF-treated HUVECs and TBP for siCOLV-treated HUVECs. n=3. (D-G) Data are presented as mean ± SD. *P < 0.05 and **P < 0.01 (D-E) LSS+HF or (F-G) LSS+siCOLV vs LSS by a ratio paired t-test.