E-TABM-360 - RNAi knock down of PLC gamma 2, CaMKII, PKC delta and Pyk2 in mouse B cell line A20 after stimulation by anti-IgG to examine the adaptation response of the BCR-dependent intracellular signaling network
Submitted on 22 October 2007, released on 1 February 2008, last updated on 2 May 2014
We examined the adaptation response of the BCR-dependent intracellular signaling network, to targeted perturbations induced through siRNA-mediated depletion of select signaling intermediates. The robustness of this network was found to derive from the fact that its constituent nodes displayed graded sensitivities to these perturbations. Importantly, this latter property also facilitated generation of perturbation-unique calibration of signal at individual nodes, with a corresponding effect on the cellular phenotypic response. Subsequently generated data-driven models revealed that the information specifying cellular phenotypic responses was not coded within variations of groups of nodes acting in a discrete manner. Rather, characteristic signaling features were selectively extracted from the majority of the constituent nodes, and then integrated to define a response-specific axis. Thus, stimulus-induced cell fate decisions are governed through a mechanism that defines the composition of signal extracted, from the overall noise provided by the assortment of signals generated at each of the nodes. The microarray experiment was performed here to identify differences in cellular responses upon various cellular perturbations in terms of gene expression profile. Cells were transfected with specific siRNAs against PLC gamma 2, CaMKII, PKC delta and Pyk2 in addition to mock siRNAs.
RNAi profiling by array, cellular modification, co-expression, compound treatment, in vitro
Capturing cell-fate decisions from the molecular signatures of a receptor-dependent signaling response. Dhiraj Kumar, Ravichandran Srikanth, Helena Ahlfors, Riitta Lahesmaa and Kanury VS Rao. Mol Syst Biol (2007)