Expression profiling of MAST. (A) Shows the protein profile of cultures induced at 2 h, 4 h and 6 h after induction with IPTG. The lysates (L) show overexpressed protein from 2 h onwards (indicated by arrow). Corresponding pellets (P) did not contain the overexpressed protein. Lysate from uninduced cell served as negative control. (B) Panel B shows the profile of the proteins eluted using a Ni-NTA column at 50, 150, 250, 300 and 500 mM concentrations of imidazole. Pure protein out in 300 and 500 mM imidazole. Lysate and the flow through fractions were loaded as controls. (C) Multi-tissue western blotting with antibody raised against MAST protein using extracts from mouse brain, kidney, ovary and testis shows a signal only in testis, indicating testis specific expression. Tubulin antibody was used as loading control. (D) Semi quantitative RT PCR using 6 tissues (kidney, brain, heart, spleen, liver from males and females, testis and ovary) shows a signal only in testis confirming the male restricted and testis specific expression of the MAST transcript. (E) Western blot analysis using testicular lysates from mouse and rat with the MAST antibody showed expression in mouse, not in rats.
Bhattacharya et al. BMC Cell Biology 2013 14:50 doi:10.1186/1471-2121-14-50