We showed that larger pieces of tissue displayed a strong gradual reduction of staining intensity from the tissue border to the center and in addition we were able to show a strong reduction of staining intensity for all tested phospho-specific antibodies beyond 2 mm in a cell pellet generated from LNT-229 glioma cells, also here we observed a stable expression without a gradient for the RPS6 and 4EBP1 total proteins. The gene discussed is RPS6; the disease is glioma.