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. 2015 Apr 2;16(4):7377–7393. doi: 10.3390/ijms16047377

Figure 5.

Figure 5

Sub-cellular localization of GCKR in seven-day old rats. (A,B) Confocal immunofluoresence micrographs with antibodies against GCKR (revealed in green), GCK (revealed in red) with nuclei counterstained with Hoechst (stained in blue) in seven-day old (A) and adult (B) rats. Overlap of the three channels is shown in Merge. White color in the merged panels denotes co-localization of GCKR and GCK in the nucleus. Slides incubated with only the secondary antibodies were used as negative controls. Images are representative of four experiments with similar results. Scale bar = 50 μm; (C) Nuclear/cytoplasmic ratio (N/C ratio) for GCKR and GCK derived from immunoflouresence. Data are presented as means ± S.D. from four individual hepatocyte preparations with 30 cells in total. *** p < 0.001 vs. 7-day old rat. (D) Representative Western blots for GCKR and GCK protein from the cytosolic (C) and nuclear (N) fractions. Abundance of GCK and GCKR in the cytosolic and nuclear fractions was normalized for comparable amounts of protein. Hsp90 and HDAC are markers for the cytoplasmic and nuclear fractions, respectively; (E) Densitometry analysis of the immunoblots in (D). Intensity of the protein band in the nuclear fraction of the adult rat is set as 100. Data are representative of four experiments with similar results.