Figure 4
Quantification of AQP5 mRNA and AQP5 protein in cancerous ovaries. A. Quantification of AQP5 mRNA in cancerous ovaries. Total RNA extracted from cancerous and normal ovaries (n = 5 animals), was DNAse-I digested and reverse transcribed. Following reverse transcription, approximately 50 ng of cDNA was used in quantitative real-time PCR using SYBR® green as the dye to quantify AQP5 mRNA and β-actin mRNA in separate reactions. Each reaction was run in triplicate per tissue sample; and the critical threshold (CT) values, averaged, normalized to that of β-actin mRNA and converted from log-linear to linear term; B. Quantification of AQP5 protein in cancerous and normal ovaries. Protein extracts from cancerous ovaries (n = 5 animals) were subjected to Western blot analysis under reducing conditions. Chicken AQP5 protein was detected by immunostaining using anti-human AQP5 antibody and chemiluminescence. Alpha-tubulin was used as loading control. Data are represented as mean ± standard error of the mean from 5 replicates. *P < 0.05; cancerous versus normal ovary.