Figure 4. Comprehensive analysis involving a variety of signal transduction molecules in transient expression of the tuberin variants.

(a) Transient expression of the tuberin variants. HEK293 cells were transiently transfected with each expression vector plasmid and subjected to the following analyses. The blots have been cropped focusing on the bands or interest; See Supplementary Fig. S3 for full-length blots. (b) Phospho-Receptor Tyrosine Kinase (RTK) Array analysis. Levels of Phospho-RTK were assessed using a horseradish peroxidase-conjugated phospho-tyrosine antibody followed by chemiluminescence detection. No significant differences were observed in the relative tyrosine phosphorylation statuses of 44 different RTKs investigated in forced transient WT- or NSM-tuberin expressing cells. (c) Reverse Phase Protein Array (RPPA) analysis. The relative concentration values of the 180 anti-phospho antibody targets are represented as a heat map. No prominent differences were observed.