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Fig. 6. Growth defect of bi-transgenic keratinocytes in vitro. (a) Image of 7-day old keratinocyte cultures from wild-type and K5tTA/TRE-KLF5 pups. Bar, 50 µm. (b) Growth curves of wild-type (
) and bi-transgenic keratinocytes (
) determined using the WST-1 reagent. The data are representative of three bi-transgenics and three wild-type cultures. (c) Addition of doxycyline to the culture prevents the growth defect; wild type (blue bars), bi-transgenic (pink bars). (d) Immunofluorescent detection and quantification of BrdU incorporation in wild-type (blue bars) and bi-transgenic (pink bars) keratinocytes showing a proliferation block in the bi-transgenics. Bar, 50 µm. In c-d the numbers represent the mean and error bars ±s.e.m. from two bi-transgenic and two wild-type cultures. To quantify BrdU incorporation at least 500 cells were counted for each culture in five fields.
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