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. 2004 Jul 12;101(29):10768–10773. doi: 10.1073/pnas.0404105101

Fig. 2.

Fig. 2.

PPI-2458-induced growth inhibition in HFLS-RA (•) and HUVEC (○), but not NHDF-Ad (▪) (A), is linked to MetAP-2 enzyme inhibition (B). (A) HFLS-RA and NHDF-Ad (8 × 103 cells) and HUVEC (8 × 103 cells) were incubated with increasing concentrations (10-12 to 10-6 M) of PPI-2458 for 7 and 4 days, respectively. For the final 24 h, 1 μCi/well of [3H]thymidine was added, and proliferation was determined by [3H]thymidine incorporation. The results are representative of at least five independent experiments. (B) The MetAP-2 assay was performed with cell lysates from HFLS-RA, NHDF-Ad, and HUVEC treated exactly as described in A, except that no [3H]thymidine was added. The amount of free MetAP-2 in each cell lysate was measured as described in Materials and Methods. Total amounts of MetAP-2 in PPI-2458-free cell lysates (=100%) were 47 pg/mg of cellular protein in HFLS-RA, 53 pg/mg in NHDF-Ad, and 7 pg/mg in HUVEC.