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Figure 1. Tropoelastin mRNA expression by HGF and HPLF. (A) Northern blotting analysis. HGF and HPLF were harvested at different times, as indicated. Total cell RNA was prepared, and 2 µg was analyzed by Northern blotting, as described in MATERIALS & METHODS. The top panel shows a representative Northern blot probed for tropoelastin mRNA expression, after which the blot was stripped and re-probed for ß-actin mRNA expression (bottom panel). (B) The bands in (A) were analyzed densitometrically, and the HGF tropoelastin mRNA to ß-actin mRNA ratios are plotted on a bar graph. The results are expressed as percentages relative to the four-week value and represent the means ± SD (standard deviation) of three independent experimental determinations. (C) Photograph of ethidium-bromide-stained 1% agarose electrophoresis gels showing the RT-PCR products of mRNAs extracted from HGF and HPLF after culture for 4 wks. A tropoelastin band from the HGF sample appearing at 1100-bp sites is present in lane 1. No detectable tropoelastin band was obtained from HPLF (lane 3). The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) bands from HGF and HPLF samples appeared at 998-bp sites in lanes 2 and 4, respectively. The molecular mass standards (lane STD) were Lambda DNA/(EcoRI and HindIII) fragments.
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