Transcriptome analysis of the Golgi stress response

We explore the transcriptional response of mammalian cells undergoing various insults to Golgi homeostasis. HEK293 cells (Flp-In T-REx 293 cells) stably containing a doxycycline-inducible Golgi-localized HaloTag2 construct (GA-HT2) were treated with the ionophore nigericin, the glycosylation inhibitor xyloside, or were induced by doxycycline and treated with the hydrophobic tag HyT36 to induce destabilization of GA-HT2. We found that while nigericin and xyloside induce global transcriptional changes, destabilization of GA-HT2 induces a Golgi-specific response. HEK293/GA-HT2 cells were treated in duplicate with nigericin, xyloside, HyT36, or the vehicle DMSO in the absence of doxycycline. In addition, cells were treated with DMSO or HyT36 in the presence of doxycyline.