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Novel luciferase-based reporter system to monitor activation of ErbB2/Her2/neu pathway noninvasively during radiotherapy.

In order to monitor activation of the UPR pathway, we investigated the expression level of GRP78, phospho-eIF2α and unconventional splicing of Xbp1 mRNA.

The induction of AMP genes depends upon signaling through the Toll and/or Imd pathway and therefore AMP expression has been used to monitor activation of these pathways during infection.

To further analyse and monitor activation of p38MAPK upon serum treatment, CYLD+/+ MEFs were serum starved for 24 hours before re-addition of serum for 30, 60 and 120 minutes.

We used primers in FlyPrimerBank to monitor activation of the JNK signaling pathway.

To monitor activation of the canonical NF- κB pathway, we analyzed I κB α phosphorylation.

We monitored activation of five protein kinases that are perturbed during viral infection and reactivation, including phospho-ERK (p-ERK), p-AKT, p-p38, p-JNK, and p-IκBα (Supplementary Table S1).

To determine whether the sensitivity of set2Δ was due to defective DNA-damage signalling, we exposed asynchronous cultures of WT and set2Δ strains to either MMS (M) or phleomycin (P) and monitored activation of γ-H2A.X (or H2AS129ph) and Rad53 (yeast homologue of mammalian Chk2).

To test whether this might also be through the ATM/APMK pathway, we monitored activation of ATM and AMPK.

We prepared alternative inputs for the eight signaling molecules and monitored activation of the integrated transcriptional reporter PFUS1-GFP (Figure 5, details in Materials and Methods).

Since activation of these stress responses could be due to a general increase in stress associated with cell death, we also monitored activation of the cytoplasmic stress response mediated by σ32 using the σ32-regulated htpG-lacZ reporter fusion.