Nuclear localization of propiece IL-1α in HeLa cells.

ABSTRACT

PURPOSE: The study aimed to examine the nuclear localization of propiece interleukin (IL)-1α (ppIL-1α) and extracellular release rates of ppIL-1α, pIL-1α, and mIL-1α. METHODS: The subcellular localization of IL-1α molecules was observed in HeLa cells transfected with green fluorescent protein (GFP)-tagged IL-1α. Extracellular release efficiency was examined using N-terminal HiBiT-tagged IL-1α. The nuclear localization status of ppIL-1α was examined by incubating ppIL-1α transfectants with 0.1% Triton X-100 solution or with complete medium on ice. RESULTS: The results indicated the diffuse cytoplasmic and nuclear localization for m and p and ppIL-1, respectively. All IL-1α forms were released from the cells even in the steady state, and the release efficiency was 25%, 13%, and 8% for mIL-1α, pIL-1α, and ppIL-1α, respectively. Under oxidative stress condition, GFP-mIL-1α was totally diminished, but weak staining of GFP-pIL-1α and GFP-ppIL-1α was detected; nuclear localization of GFP-ppIL-1α was completely abolished by 0.1% Triton X-100 treatment, however, it remained in the nucleus after culture in complete medium on ice. CONCLUSION: The results of this study showed that ppIL-1α was localized in the nucleus and released extracellularly even in the steady state. Moreover, its cellular localization is not firm, and it is presumed to be floating in the nucleoplasm.