Skip to main content
Fig. 2 | Molecular Medicine

Fig. 2

From: Altered Procollagen mRNA Expression during the Progression of Avian Scleroderma

Fig. 2

RPA of total RNA from comb tissue of UCD-200 and UCD-058 chickens

Two micrograms of RNA was hybridized with two sets of Dig-UTP-labeled probes (set 1: antisense probes for α1(I), α2(I), α1(II) as a negative control, and α1(VI) procollagen and GAPDH; set 2: antisense probes for α1(III), α2(VI), and α3(VI) procollagen and GAPDH). Dig-UTP-labeled RNA was detected by chemiluminescence. Tissue samples were from UCD-200 without clinical symptoms (0), erythema (1 +), edema (2 +), beginning necrosis (3 +) of the comb, and from the comb of 8-month-old (8m) UCD-200 chickens in the late chronic stage of the disease, as well as tissue samples from 11-day-old (11d) and 8-month-old UCD-058 controls. (A) RPA blot: Ambion’s Century-size-marker (SM) was used to estimate band size. Arrows indicate the two mRNA variants of α2(I) procollagen. In some lanes on the presented blots the lower α2(I) and the α3(VI) band are not visible since we had to find an exposure time that would show both strong bands and weak bands on the same blot. (B) Quantification: For densitometrical analysis, several films were exposed for each blot at varying times and used to calculate the relative signal abundancies and ratios of strong and weak bands via an exponential fitting curve to account for the nonlinear blackening of X-ray films. Signal intensities of the different procollagen transcripts were added and taken as 100%. The UCD-200 groups were statistically compared with age-matched UCD-058 controls by using a Mann-Whitney-Wilcoxon-U test; *p < 0.05; **p < 0.025; ***p < 0.01.

Back to article page