Figure 2.

Diced-siRNA screen of the human signaling proteome identifies regulators of transferrin uptake. (a) Selected d-siRNAs include proteins with known signaling related domains (as named and annotated in the NCBI CDD database of 20 July 2005 [29]. Related domains were pooled in the same category bin). Transmembrane receptors and transcription factors were excluded. (b) Examples of signaling pathways and numbers of their components represented in the selected signaling proteome set. (c) Measured transferrin uptake values (F) from the human signaling proteome screen. Data sorted in ascending order. (d) Method to calculate hit probabilities by CAsH analysis. The histogram of the screen data (light blue) is compared to the stochastic noise distribution (red). CAsH values correspond to the estimated fraction of hits for each F score. (e) Regions within mRNAs selected to design the two independent d-siRNA pools. Seq1 corresponds to the region chosen to synthesize the initial batches of d-siRNAs, Seq2 to the region chosen to synthesize the second, independent batch. (f) Transferrin uptake regulators identified by two independent d-siRNA pools. (g) Genes increasing or decreasing transferrin uptake in HeLa cells.