10.6084/m9.figshare.5126974.v1 Alekseev O. Alekseev O. Limonnik V. Limonnik V. Donovan K. Donovan K. Azizkhan-Clifford J. Azizkhan-Clifford J. Supplementary Material for: Activation of Checkpoint Kinase 2 Is Critical for Herpes Simplex Virus Type 1 Replication in Corneal Epithelium Karger Publishers 2014 Herpes simplex virus type 1 Keratitis Checkpoint kinase 2 Corneal epithelium Explant cornea DNA damage response Small-molecule inhibitor 2014-12-19 00:00:00 Dataset https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Activation_of_Checkpoint_Kinase_2_Is_Critical_for_Herpes_Simplex_Virus_Type_1_Replication_in_Corneal_Epithelium/5126974 <b><i>Background/Aims:</i></b> Herpes simplex virus (HSV) type I keratitis remains a leading cause of corneal morbidity, despite the availability of effective antiviral drugs. Improved understanding of virus-host interactions at the level of the host DNA damage response (DDR), a known factor in the development of HSV-1 keratitis, may shed light on potential new therapeutic targets. This report examines the role of checkpoint kinase 2 (Chk2), a DDR mediator protein, in corneal epithelial HSV-1 infection. <b><i>Methods:</i></b> A small-molecule inhibitor of Chk2 (Chk2 inhibitor II) was applied to HSV-1-infected cultured human corneal epithelial cells (hTCEpi and HCE) as well as to explanted and organotypically cultured human and rabbit corneas. Infection levels were assessed by plaque assay and real-time PCR. RNAi-mediated depletion of Chk2 was performed to confirm the effect of the inhibitor. <b><i>Results:</i></b> Inhibition of the Chk2 kinase activity greatly suppresses the cytopathic effect, genome replication and infectious progeny production in vitro and ex vivo. <b><i>Conclusion:</i></b> This report demonstrates the critical role of Chk2 kinase in the establishment of HSV-1 corneal epithelial infection. These data contribute to our understanding of herpesvirus-host interactions and underscore the significance of DDR activation in HSV-1 keratitis.