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. 2018 Apr 27;9(32):22586–22604. doi: 10.18632/oncotarget.25187

Figure 3. LRP5 depletion induces apoptosis more robustly than LRP6 depletion.

Figure 3

MDA-MB-468 and HCC38 cells were transfected with one of two different siRNAs against LRP5 (#2 and #4, red) or LRP6 (#7 and #8, blue) or with a control siRNA (black). (A) The activation of executioner caspases 3/7 was assessed 72 to 144 h after transfection, in a Caspase-Glo®3/7 luminescence assay. The results are presented as caspase3/7 activity normalized against the caspase activity in control siRNA-transfected cells. The data shown are the means ± SD for three independent experiments. In some conditions, caspase activity levels were lower than those in the control, because the results obtained in this assay depend in part on the number of cells. (B) The activation of caspases 3/7 in the presence or absence of the pan-caspase inhibitor Z-VAD-FMK was assessed 120 hours after the depletion of LRP5, as in Figure 3A. The data shown are means ± SD for 2 independent experiments performed in triplicate. (C) PARP cleavage (c-PARP) and the activation of caspases 3, 7 and 8 were analyzed by western blotting 96 hours after transfection with control (ctrl), LRP5 or LRP6 siRNAs. One representative experiment of the five performed, all of which gave similar results, is shown. Actin and GAPDH were used as loading controls. ***P<0.001 (i.e. an increase relative to control conditions).