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Fig. 4. GAP-dependent Rac inactivation sensitizes mammary acini to induction of death. (A) Representative immunoblot showing non-detectable amounts of hemagglutinin (HA) expressed in vector control MECs and high levels in MECs expressing an exogenous HA-tagged 
2-chimerin. (B) Representative immunoblot showing GTP-Rac, Rac and E-cadherin levels in vector control MECs in comparison with 2-chimerin-expressing MECs. (C) Average relative specific-activity of GTP-loaded Rac illustrating reduced Rac activity in MEC-expressing exogenous 2-chimerin in comparison with vector control MECs. (D) Bar graph illustrating increased percentage of apoptotic cells induced by treatment with Trail (1 µg/ml) in 3D rBM differentiated MECs with reduced Rac activity mediated by expression of either the Rac GAP 2-chimerin or dominant-negative N17Rac. The percentage apoptosis was calculated by scoring the number of activated caspase-3-positive MECs divided by the total number of MECs 24 hours following treatment with Trail. Similar results were obtained following treatment with chemotherapeutic agents (data not shown). (E) (top) Phase-contrast and (bottom) immunofluorescence images of 3D poly-HEMA rBM-generated vector control MEC colonies (left) and MEC colonies expressing HA-tagged 2-chimerin (right; red) illustrating robust and uniform expression of the transgene 48 hours following adenoviral infection. Bar, 50 µm. Arrows indicate the representative phenotype of acini morphology. Bar, 50 µm. Results are the mean ± s.e.m. of three separate experiments. *P
0.05; **P0.01.
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