Figure 4

Loss of TRIP-1 in human lung fetal fibroblasts leads to decreased apoptosis in a collagen matrix. A: TUNEL staining in HLF-F cells transfected with control siRNA (left) or TRIP-1 siRNA (right). Cells were transfected, and 2 days later embedded in a type I collagen matrix and allowed to contract matrix for 5 days, then cells were recovered from the gel and staining performed as described in Methods. Staining in green shows positive TUNEL, while blue corresponds to DAPI; B: Quantification of % positive TUNEL stained cells (apoptosis) based on analysis of pictures like the ones shown in A. Experiment was repeated three times, one experiment is shown. Means ± SE shown (**P < 0.05, n = 3); C: control siRNA or TRIP-1 siRNA-transfected cells were embedded in collagen matrix, gels were collected after 24 or 48 hours and lysed directly in sample buffer (Laemli); western-blot for cleaved caspase 3 is shown. Actin is shown as a protein loading control; one representative Western-blot experiment is shown, out of three performed.