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. 2012 Mar 16;19(9):1514–1524. doi: 10.1038/cdd.2012.28

Figure 2.

Figure 2

Actin depolymerization by villin is required for its anti-apoptotic function. MDCK Tet-Off cells expressing full-length and mutant villin proteins were cultured in the presence (VIL/NULL) or absence of doxycycline and treated with CPT (20 μM, 0–5 h). (a) VIL/NULL- and villin-expressing cells were visualized by phase contrast microscopy and stained with Hoechst 33258 to identify apoptotic cells. This is representative of six other experiments with similar data. Bar, 50 μm. PI permeability was analyzed by flow cytometry as a quantitative measure of cell viability. There was a significant increase in PI positive VIL/NULL, VIL/ΔPB2 and VIL/RKR cells in response to CPT treatment (*P<0.01, n=9). (b) Caspase-3 activity was measured in MDCK Tet-Off VIL/NULL- and villin-expressing (wild-type and mutant) cells treated without or with CPT (20 μM) for 0–6 h. Caspase-3 activity was significantly inhibited in VIL/WT, VIL/ΔPB1, VIL/ΔPB5 and VIL/R138A cells as early as 4 h post treatment (*P<0.01, n=6)