Figure 5. Effects of JK-31 on proliferation and cell cycle status of primary human endothelial cells and human breast cancer cells.

(A) HUVECs and (B) MCF-7 cells were treated with DMSO, JK-31 (0.1, 1, 10, 50 µM), vatalanib (10 µM) or bohemine (10 µM) in full growth medium for 16 h followed by labeling with bromodeoxyuridine (BrdU; 10 µM) for 2 h and subsequent ELISA. Absorbance readings at OD₄₅₀ were normalized and expressed relative to DMSO control. Error bars represent ± SEM (n = 4; ***p<0.001). (C) HUVECs and (D) MCF-7 cells were treated with nocodazole (200 nM), bohemine (10 µM), sunitinib (100 nM) or JK-31 (0, 0.1, 1, 10, 50 µM) for 48 h. Total cell lysates were prepared and immunoblotting carried out as described in Experimental Section. Membranes were probed for phosphorylated and total CDK1 in addition to cyclin A, B and D1 and a loading control (α-tubulin). Representative immunoblots of three independent experiments shown.