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. Author manuscript; available in PMC: 2019 Feb 3.
Published in final edited form as: Biochim Biophys Acta. 2018 Feb 3;1861(2):72–79. doi: 10.1016/j.bbagrm.2018.01.002

Fig. 1.

Fig. 1

Temporal analysis of ATF3 transcription activity during the AAR and UPR. HepG2 cells were cultured in DMEM (Control) ± 2.5mM HisOH or±50 nM Tg for 0–24 h. Total RNA was isolated at the indicated times and RT-qPCR was performed to analyze the ATF3 transcriptional activity as measured by hnRNA (A) and steady-state mRNA levels of ATF3 (B). To verify that the decreased ATF3 transcriptional activity in DMEM+HisOH was not the result of HisOH degradation, transcriptional activity was measured in HepG2 cells in which the media in one half of the cells was supplemented with fresh 2.5mM HisOH at the 4 h time period (C) or in cells incubated in DMEM±histidine for 0–24 h (D). In all panels, GAPDH mRNA, which is not affected by the AAR or UPR, was used as an internal control. The results shown are the means±S.D. of at least three replicates. Each experiment was repeated to ensure reproducibility between batches of cells.