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Fig. 2. Expression of the Xl MMP-9TH gene is induced during spontaneous metamorphosis and in a T3-treated XLT-15 cultured cell line. (A) The developmental expression of Xl MMP-9TH mRNA in tail, hindlimb, intestine and central nervous system. A Xl MMP-9TH probe was hybridized to 5 µg of total cellular RNA isolated from tail (a), hindlimb (b), intestine (c) and central nervous system (d) of stage 56–63 tadpoles. These blotting filters contain RNA of stage 58 tail for the comparison. Arrowheads indicate the positions of 18S and 28S rRNA. (B) The T3 dose–response of Xl MMP-9TH mRNA upregulation. A Xl MMP-9TH probe was hybridized to 5 µg of total cellular RNA isolated from XLT-15 cells treated with the indicated concentration of T3 for 24 h. (C) The time course of Xl MMP-9TH mRNA upregulation by T3. A Xl MMP-9TH probe was hybridized to 5 µg of total cellular RNA isolated from XLT-15 cells treated with 10 nm T3 for the indicated times. (D) The effect of protein synthesis inhibition on Xl MMP-9TH mRNA upregulation by T3. A Xl MMP-9TH probe was hybridized to 5 µg of total cellular RNA isolated from XLT-15 cells treated with or without 10 nm T3 for 8 h in the presence or absence of a protein synthesis inhibitor, 10 µg/mL cycloheximide. Control hybridization of the blots with the Xenopus laevis elongation factor-1α probe (EF) (Krieg et al. 1989) is shown below to standardize the amounts of RNA. |
