The Dual-Light System Kit (Applied Biosystems) was used to measure the β-galactosidase activity in E14.5 embryo tissue homogenate. Whole tail and limbs of E14.5 embryos (n = 5 forTIMP3-Tg heterozygotes and n = 7 for [-1A]TIMP3-Tg heterozygotes) were homogenized in lysis buffer using a micropestle (Eppendorf). The homogenate was centrifuged, and the clear supernatant obtained was subjected to analysis as per the manufacturer’s instructions. Light output was quantified as relative light units using a luminometer (Berthold Technologies). To normalize enzyme activities, the total protein concentration of lysates was determined using the BCA Protein Assay Kit (Pierce).
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