Secretion mode and subcellular localization of human interferon-beta exogenously expressed in porcine renal epithelial LLC-PK1 cells.

We have investigated the secretion polarity of human interferon-beta (HuIFN-beta) exogenously expressed in the porcine renal proximal tubule cell line, LLC-PK1. In these cells, stably expressed HuIFN-beta was secreted to the apical and basolateral sides. However, when transiently expressed by apical lipofection, HuIFN-beta was secreted to both cell sides, while basolateral-preferential secretion was seen for basal transfection.

Simultaneous detection of DsRed2-tagged and EGFP-tagged human beta-interferons in the same single cells.

The red fluorescent protein DsRed2 is a useful fusion tag for various proteins, together with the enhanced green fluorescent protein (EGFP). These chromoproteins have spectral properties that allow simultaneous distinctive detection of tagged proteins in the same single cells by dual color imaging. We used them for tagging a secretory protein, human interferon-beta (IFN-beta).

Subcellular trafficking of exogenously expressed interferon-beta in Madin-Darby canine kidney cells.

We have recently demonstrated that when IFN-beta was exogenously expressed in epithelial cells, transiently expressed IFN-beta was predominantly secreted from the cell side to which the transfection was performed, while stably expressed one was almost equally secreted to the apical and basolateral sides. In the present study, we analyzed the subcellular transport of IFN-beta using confocal imaging with green fluorescent protein (GFP)-tagged IFN-beta in Madin-Darby canine kidney (MDCK) cells. Stably expressed and transiently expressed human IFN-beta (HuIFN-beta)-GFPs were seen in upper regions of the nucleus.