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. 2018 Jan;188(1):111–124. doi: 10.1016/j.ajpath.2017.09.009

Figure 4.

Figure 4

Dual immunofluorescence staining for the co-localization of mechanistic target of rapamycin (mTOR) and casein kinase (CSNK)-2β. Human hepatocellular carcinoma cells were stained with anti-mouse mTOR and anti-rabbit CSNK-2β antibodies. Corresponding secondary antibodies were Alexa anti-mouse 568 and Alexa anti-rabbit 660, respectively. Images were captured via confocal microscopy. A: mTOR (red) is present throughout the cell but most abundant in the nucleus. B: CSNK-2β (green) is found throughout the cell. C: Merged channel image shows co-localization (yellow). D: Because DAPI staining obscured the depiction of yellow co-localization, dual-immunofluorescence staining is visualized without DAPI. E: To determine the cytosolic co-localization of mTOR and CSNK-2β, we subtracted the positive DAPI staining area from the whole cell image. F: With the use of the positive DAPI stain, nuclear co-localization of mTOR and CSNK-2β is observed. Scale bars: 20 μm.