Eight Stuff You Did Not Understand Or Know Concerning BIBW2992
A targeting vector containing the transgene shown in Fig.?1A was generated and used to produce founders by standard SWAP70 pronuclear injection in FVB/N background; 3 founder lines were obtained (founder #8 a female and founders #10 and #16, males). Southern blotting (Fig.?1B), showed that the signal for the transgene was greater than the RAR�� signal. Hemizygous lines were established for all 3 founders and transgenic males were mated with wild type females. As shown for lines #8 and #16 (Fig.?1C), the transgene was highly expressed in adipose tissue, including the forth mammary fat pad, abdominal white adipose tissue (WAT) and interscapular brown adipose tissue (BAT). Low level of expression in other organs was, likely, due to fat contamination. Staining of the transgenic mammary for HA tag (Fig.?1D, right panel), showed that its expression was limited to adipose tissue and excluded from the mammary epithelium. As expected, the wt-mammary was negative for HA (Fig.?1D, left panel). All 3 lines of the transgenic mice were visibly smaller, had coarser fur and their body weight, as well as the weight of their selleck mammary fat pads, was ~?10% lower than in the wild type, even at week 10. Other tissues, such as kidneys, heart and brain did not differ in weight (results not shown). The transgenic and the wt-type mammary fat-pad adipocytes had similarly appearing fat-filled cells with large, Oil Red O stained vacuoles suggesting that the decrease in weight is due to reduced number and not due to inhibited differentiation of adipocytes. Whole mounts of BIBW2992 price mammary glands from control and Tg-mice at 6, 9 and 12?weeks after birth were examined. As shown in Fig.?2A, the epithelial duct outgrowth in the Tg gland was drastically reduced at 6?weeks, and remained smaller even at 12?weeks when the wt epithelial ducts reached the end of the fat pad. Very similar changes were observed in the 2 additional Tg-lines (results not shown). These changes were quantified by measuring the ductal length from the nipple area to the tip of the longest duct and the length of the fat pad from the nipple to the edge of the fat pad. As shown in Fig.?2B, the ducts in the transgenic glands were persistently (weeks 6 to 16) and significantly (p?
