Knockdown of ZNF24 leads to decreased MMP-2 levels without affecting levels of major MMP-2 activity regulators. A, C, and E) HMVEC-D cells were transfected with control siRNA or siZNF24, and mRNA levels of TIMP1 (A), TIMP2 (C), and MMP14 (E) were analyzed by qPCR. ***P < 0.001; N.S., not significant. Values in all graphs represent mean ± sd from 3 independent experiments. B and D) HMVEC-D cells were transfected with control siRNA or siZNF24, and protein levels of TIMP-1 (B) and TIMP-2 (D) in the conditioned media were analyzed by ELISA. ***P < 0.001; N.S., not significant. F) HMVEC-D cells were transfected with control siRNA or siZNF24. Protein samples from 3 independent experiments were analyzed by immunoblotting and quantified by densitometry using Adobe Photoshop CS5. *P < 0.05; N.S., not significant. G) HMVEC-D cells were transfected with control siRNA or siZNF24. Conditioned media treated with or without APMA were analyzed by substrate gel electrophoresis for MMP-2 activity (lanes 3–6). Purified MMP-2 treated with or without APMA was used as control samples (lanes 1 and 2). This experiment was conducted 3 times, and a representative experiment is shown.