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. 2017 Feb 28;108(2):208–215. doi: 10.1111/cas.13133

Figure 3.

Figure 3

Small interfering RNA‐based screening of Cullins (CULs) responsible for regulating vascular endothelial (VE)‐cadherin. (a) HUVECs were transfected with MISSION siRNAs (Sigma‐Aldrich) against control (CONT), CUL1,CUL2,CUL3,CUL4A, and CUL5, followed by culturing for 72 h. The proteins were extracted and separated using 10% SDS–polyacrylamide gels. VE‐cadherin proteins were then detected by Western blot analysis. β‐Actin was used as a loading control. (b) Quantification of VE‐cadherin protein levels in CONT,CUL1,CUL2,CUL3,CUL4A, and CUL5 knockdown cells was carried out using ImageJ software. ***P < 0.001. (c) VE‐cadherin mRNA levels were quantified by quantitative RT‐PCR in CONT,CUL1,CUL2,CUL3,CUL4A, or CUL5 siRNA‐transfected HUVECs, and normalized to GAPDH mRNA levels.