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. 2010 Aug 2;285(42):32638–32646. doi: 10.1074/jbc.M110.105544

FIGURE 4.

FIGURE 4.

Id1 knockdown in HMEC-1 cells caused disruption of tube formation on Matrigel. A and B, HMEC-1 cells were plated into six-well plates for 24 h before transfection with Id1 shRNA for 30 h. Both RT-PCR and Western blotting were then used to detect Id1 expression. Id1 shRNA could down-regulate Id1 expression at mRNA level (A) and protein level (B). C, the tube formation of HMEC-1 cells on Matrigel was disrupted (panel c) after Id1 shRNA transfection, compared with the blank (panel a) and sham control (panel b). HMEC-1 cells (2.5 × 105 cells/well) were seeded into a six-well plate for 24 h before transfection. The shRNA plasmids were transfected into the cells three times 24 h apart using X-fect polymer (Clontech). The cells were used in the tube formation assay as described under “Experimental Procedures” 24 h after the last transfection. Photos were taken after another 12 h of incubation. The results are representative of three experiments.