Detection and functional analysis of TIMP-1. (A) Western blot analysis of unconcentrated TIMP-1 in conditioned media of various BL lines: lane 1, rTIMP-1 (300 ng) positive control; lane 2, JD38; lane 3, ST846; lane 4, Daudi; lane 5, AG876; lane 6, PA682; lane 7, DW6. JD38 cells were transfected with human timp-1 or emptyLXSN vector as described in the Materials and Methods. (B) Western blot analysis of TIMP-1 by TIMP-1–JD38 cell clones: lane 1, rTIMP-1 (300 ng) positive control; lane 2, parental JD38 cells; lane 3, TIMP-1–JD38 cell clone 20; lane 4, TIMP-1–JD38 cell clone 24; lane 5, LXSN-JD38 cells. (C) Reverse zymogram of TIMP-1 in the concentrated conditioned media of transfected JD38 cells: lane 1, LXSN-JD38 cells; lane 2, TIMP-1–JD38 cell clone 20; lane 3, TIMP-1–JD38 cell clone 24; lane 4, positive control HT1080 cells. Representative results of three independent analyses.