Figure 5
Highest expression of Smoc -1 in liver. RT-PCR showed expression in liver only but Southern hybridization detected reduced signals in testis, ovary and spleen (A) whereas control RT-PCR with β-actin showed almost equal intensity signal in each tissue (B). Quantitative expression of Smoc-1 based on Real Time PCR confirmed maximum expression (163–364 folds) in liver in five different animals compared to that in lung used as the calibrator (C). Note the buffalo spermatozoa from four different animals (BS1-4) also showed 4–7 folds transcripts in four different animals, similar to that in testis. Relative quantitation also demonstrated higher expression (1.2–3.5 times) of variant-02 compared to that of -01 in each of the tissues examined (D). Western blotting with anti-SySmoc 1-pAb (E) and anti-β-actin-mAb as positive control (F) substantiated the highest expression of Smoc-1 protein in liver. The quantitative expression carried out using cDNA isolated from blood lymphocytes of different age group of animals is shown in (G). Note the markedly increased expression of Smoc-1 in animals 10 months and beyond. Relative quantitation showing higher expression of variant-01 in the animal up to ~6 months of age and that of variant-02 in the animals of age 6 months and beyond (H).