The heterogeneous nuclear ribonucleoproteins (hnRNPs) are a family of RNA-binding proteins with important roles in different aspects of nucleic acid metabolism, as well as in the regulation of various cell signalling pathways. Herpes simplex virus 1 (HSV-1) is an important and widespread human pathogen that in most cases causes asimptomatic or mild disease such as oral or facial lesions, but can sometimes cause more serious conditions, like encephalitis and meningitis. Previous experiments have shown that hnRNPs interact with components of various viruses, including HSV-1, and that virus infection causes changes in subcellular localization, but the precise molecular mechanisms of these interactions are yet not understood. In this study, using immunofluorescence and Western blot, we aimed to test whether the localization and expression levels of hnRNPs change during HSV-1 infection, with special attention to the multifunctional hnRNP K protein. We also investigated changes in subcellular localization and expression levels in cells treated with pharmacological activators of type I interferon response: IFN-β, LPS and poly I:C, as well as with autophagy inducer, rapamycin and activator of apoptosis, staurosporine. HSV-1 infection caused hnRNP K translocation from the nucleus to the cytoplasm, which was not the case when cells were treated pharmacologically. Expression levels of hnRNPs stayed unchanged during the course of HSV-1 infection, but pharmacological treatments caused either a decrease or increase in expression of hnRNP K, hnRNP E2 and hnRNP U. By comparing the localization and expression of hnRNPs during HSV1 infection and farmacological treatments, this study contributes to better understanding of how these proteins participate in the complex interaction between virus and host organisms.